LYPLA2

cell_type

HeLa

experimental_model

Enzyme perturbation and pulse-chase

limitations

Not removal of the catalytic acyl intermediate.

organism

human

Selenium: literature corrections and mechanism additions · lines 894–904

Metabolic Ledger literature curation, 17 September 2026; primary papers linked individually · supports · Enzyme perturbation and pulse-chase · secondary_verified · secondary_verified

## apt2-depalmitoylates-zdhhc6 APT2 can remove ZDHHC6's regulatory lipid groups. APT2/LYPLA2 removes regulatory palmitoylation from ZDHHC6, with rapid turnover involving C328. Organism: human Cell type: HeLa Experimental model: Enzyme perturbation and pulse-chase Limitations: Not removal of the catalytic acyl intermediate. Primary reference: [Identification and dynamics of the human ZDHHC16-ZDHHC6 palmitoylation cascade](https://elifesciences.org/articles/27826)

experimental_model

Recombinant human APT2 SPR; supporting cellular CETSA in NOZ/GBC-SD

exposure

SPR binding concentration series; cellular CETSA used 25 micromolar solasonine. Binding does not specify catalytic inhibition.

limitations

Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

organism

Human

primary_locator

Figure 5J and Methods 2.20; CETSA Figure 5F-G.

primary_references

https://doi.org/10.1002/ptr.70245

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 60–66

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Recombinant human APT2 SPR; supporting cellular CETSA in NOZ/GBC-SD · source_derived_draft · unverified_draft

Surface plasmon resonance reported solasonine binding to recombinant APT2, KD 2.982 micromolar. primary_references: https://doi.org/10.1002/ptr.70245 primary_locator: Figure 5J and Methods 2.20; CETSA Figure 5F-G. organism: Human experimental_model: Recombinant human APT2 SPR; supporting cellular CETSA in NOZ/GBC-SD exposure: SPR binding concentration series; cellular CETSA used 25 micromolar solasonine. Binding does not specify catalytic inhibition. limitations: Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

evidence_cache

artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a

experimental_model

Human Huh-7 and HEK293 cell lysates; endogenous or FLAG-tagged human APT2

exposure

Bead pulldown; free-sulforaphane competition at 100 micromolar for 30 minutes at 4 C. C56S and C2S constructs compared.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. Mutagenesis supports C56-dependent engagement; no binding affinity or substrate-specific catalytic inhibition constant was established.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figures 1 and 3, Results 3.1 and 3.3

primary_references

https://doi.org/10.1016/j.bbrc.2024.150244

source_access

Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 8–16

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human Huh-7 and HEK293 cell lysates; endogenous or FLAG-tagged human APT2 · source_derived_draft · unverified_draft

Sulforaphane-affinity beads captured APT2 from Huh-7 or transfected HEK293 lysates; C56S mutation attenuated binding in the reported assays. primary_references: https://doi.org/10.1016/j.bbrc.2024.150244 primary_locator: Figures 1 and 3, Results 3.1 and 3.3 source_access: Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a experimental_model: Human Huh-7 and HEK293 cell lysates; endogenous or FLAG-tagged human APT2 organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: Bead pulldown; free-sulforaphane competition at 100 micromolar for 30 minutes at 4 C. C56S and C2S constructs compared. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. Mutagenesis supports C56-dependent engagement; no binding affinity or substrate-specific catalytic inhibition constant was established.

experimental_model

Human HEK293T with expressed APT2 and Flag-STAT3

exposure

Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved.

limitations

C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

organism

Human

primary_locator

Figure 2d; Extended Data 7b-c.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

APT2-STAT3 cycling and a competing sulforaphane target route · lines 51–57

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T with expressed APT2 and Flag-STAT3 · source_derived_draft · unverified_draft

APT2-C2S expression did not reproduce the wild-type reduction in STAT3 palmitoylation. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2d; Extended Data 7b-c. organism: Human experimental_model: Human HEK293T with expressed APT2 and Flag-STAT3 exposure: Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved. limitations: C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human HEK293T subcellular fractionation

exposure

LYPLA2 siRNA; exact dose/duration for this panel unresolved.

limitations

Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

organism

Human

primary_locator

Figure 2g; APT2 depalmitoylates p-STAT3.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2-STAT3 cycling and a competing sulforaphane target route · lines 24–30

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T subcellular fractionation · source_derived_draft · unverified_draft

APT2 knockdown reduced nuclear p-STAT3 in the HEK293T fractionation experiment. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2g; APT2 depalmitoylates p-STAT3. organism: Human experimental_model: Human HEK293T subcellular fractionation exposure: LYPLA2 siRNA; exact dose/duration for this panel unresolved. limitations: Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human HEK293T

exposure

LYPLA2 siRNA; exact exposure unresolved.

limitations

Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

organism

Human

primary_locator

Extended Data 7e-f and Results: APT2 depalmitoylates p-STAT3.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2-STAT3 cycling and a competing sulforaphane target route · lines 33–39

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T · source_derived_draft · unverified_draft

APT2 knockdown reduced STAT3 transcriptional activity in the reported HEK293T experiments. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Extended Data 7e-f and Results: APT2 depalmitoylates p-STAT3. organism: Human experimental_model: Human HEK293T exposure: LYPLA2 siRNA; exact exposure unresolved. limitations: Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure.

exposure

LYPLA2 siRNA before Alk14 labeling; Figure 2f states 36 hours before labeling; exact siRNA amount unresolved.

limitations

Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

organism

Human

primary_locator

Figure 2f; Extended Data 7d.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2-STAT3 cycling and a competing sulforaphane target route · lines 15–21

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure. · source_derived_draft · unverified_draft

LYPLA2 siRNA increased STAT3 palmitoylation in the reported reconstituted HEK293T experiment. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2f; Extended Data 7d. organism: Human experimental_model: Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure. exposure: LYPLA2 siRNA before Alk14 labeling; Figure 2f states 36 hours before labeling; exact siRNA amount unresolved. limitations: Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

experimental_model

Human HEK293T with expressed APT2 and Flag-STAT3

exposure

Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved.

limitations

C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

organism

Human

primary_locator

Figure 2d; Extended Data 7b-c.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

APT2-STAT3 cycling and a competing sulforaphane target route · lines 60–66

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T with expressed APT2 and Flag-STAT3 · source_derived_draft · unverified_draft

APT2-S122A expression did not reproduce the wild-type reduction in STAT3 palmitoylation. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2d; Extended Data 7b-c. organism: Human experimental_model: Human HEK293T with expressed APT2 and Flag-STAT3 exposure: Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved. limitations: C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

experimental_model

Human HEK293T with expressed APT2 and Flag-STAT3

exposure

Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved.

limitations

C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

organism

Human

primary_locator

Figure 2d; Extended Data 7b-c.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

APT2-STAT3 cycling and a competing sulforaphane target route · lines 42–48

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T with expressed APT2 and Flag-STAT3 · source_derived_draft · unverified_draft

Wild-type APT2 expression reduced STAT3 palmitoylation. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2d; Extended Data 7b-c. organism: Human experimental_model: Human HEK293T with expressed APT2 and Flag-STAT3 exposure: Expression constructs; 50 micromolar Alk14 labeling for 5 hours; expression duration unresolved. limitations: C2S disrupts the APT2 palmitoylation site; S122A alters its catalytic site. They are not interchangeable controls. Null results do not establish equivalence. GPX4 was not an endpoint.

experimental_model

Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure.

exposure

20 micromolar ML349 for 36 hours before Alk14 labeling; labeling methods specify 50 micromolar Alk14 for 5 hours.

limitations

Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

organism

Human

primary_locator

Figure 2f; Extended Data 7d; APT2 depalmitoylates p-STAT3.

primary_references

https://doi.org/10.1038/s41586-020-2799-2

APT2-STAT3 cycling and a competing sulforaphane target route · lines 6–12

Selected primary observations: 10.1038/s41586-020-2799-2 and 10.1038/s41388-020-1335-z. · supports · Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure. · source_derived_draft · unverified_draft

ML349 increased STAT3 palmitoylation in the reported reconstituted HEK293T experiment. primary_references: https://doi.org/10.1038/s41586-020-2799-2 primary_locator: Figure 2f; Extended Data 7d; APT2 depalmitoylates p-STAT3. organism: Human experimental_model: Human HEK293T; DHHC7 knockout reconstituted with HA-DHHC7 and Flag-STAT3; DHHC construct species is not established as human for this figure. exposure: 20 micromolar ML349 for 36 hours before Alk14 labeling; labeling methods specify 50 micromolar Alk14 for 5 hours. limitations: Reported cellular result, not a GPX4 time-course experiment. Overexpressed DHHC constructs in much of the study are mouse proteins; human constructs are explicitly identified only where confirmed. APT2 depletion, selective inhibitor exposure and sulforaphane are different interventions.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 33–39

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

C2S APT2 re-expression did not reproduce wild-type restoration of RSL3 susceptibility in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 24–30

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

C2S APT2 re-expression did not reproduce the wild-type reduction in GPX4 protein in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 51–57

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

S122A APT2 re-expression did not reproduce wild-type restoration of RSL3 susceptibility in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 42–48

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

S122A APT2 re-expression did not reproduce the wild-type reduction in GPX4 protein in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 15–21

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

Wild-type APT2 re-expression restored RSL3 susceptibility in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs

exposure

HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here.

limitations

C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

organism

Human

primary_locator

Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis.

primary_references

https://doi.org/10.1038/s41467-025-56344-5

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 6–12

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs · source_derived_draft · unverified_draft

Wild-type APT2 re-expression lowered GPX4 protein in APT2-depleted A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6i-j; Results: Inhibition of APT2 suppresses ferroptosis. organism: Human experimental_model: Human A375 melanoma cells with APT2 shRNA and lentiviral rescue constructs exposure: HA-tagged rescue constructs; Figure 6j: 48-hour RSL3 challenge, n=3 independent experiments. Exact RSL3 concentrations follow the figure dose response and are not inferred here. limitations: C2S affects APT2 palmitoylation/localization and is not identical to the S122A catalytic-site perturbation. Null comparisons do not prove equivalence. No sulforaphane or solasonine tested in this rescue experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

experimental_model

Human NOZ and GBC-SD gallbladder cancer cells

exposure

Methods 2.2 describes 48-hour cellular drug treatments with refresh at 24 hours; exact concentration must be checked per figure and is not inferred from another assay.

limitations

Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

organism

Human

primary_locator

Figure 6E; Supplementary Figure S5A; Results 3.6.

primary_references

https://doi.org/10.1002/ptr.70245

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 159–165

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human NOZ and GBC-SD gallbladder cancer cells · source_derived_draft · unverified_draft

Solasonine did not increase STAT3 palmitoylation in the reported APT2-D13A/L11A mutant setting. primary_references: https://doi.org/10.1002/ptr.70245 primary_locator: Figure 6E; Supplementary Figure S5A; Results 3.6. organism: Human experimental_model: Human NOZ and GBC-SD gallbladder cancer cells exposure: Methods 2.2 describes 48-hour cellular drug treatments with refresh at 24 hours; exact concentration must be checked per figure and is not inferred from another assay. limitations: Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

experimental_model

Human NOZ and GBC-SD gallbladder cancer cells

exposure

Cell-supernatant ELISA; 0-50 ng/mL standards, anti-LYPLA2 plates, LYPLA2-HRP conjugate, H2O2/TMB chromogen, 450 nm readout with 630 nm reference. Cellular drug duration described as 48 hours.

limitations

Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

organism

Human

primary_locator

Figure 5C versus Methods 2.17 ELISA Assay.

primary_references

https://doi.org/10.1002/ptr.70245

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 105–111

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human NOZ and GBC-SD gallbladder cancer cells · source_derived_draft · unverified_draft

The study reports a reduced LYPLA2 ELISA signal as reduced APT2 activity; the described assay does not establish catalytic turnover. primary_references: https://doi.org/10.1002/ptr.70245 primary_locator: Figure 5C versus Methods 2.17 ELISA Assay. organism: Human experimental_model: Human NOZ and GBC-SD gallbladder cancer cells exposure: Cell-supernatant ELISA; 0-50 ng/mL standards, anti-LYPLA2 plates, LYPLA2-HRP conjugate, H2O2/TMB chromogen, 450 nm readout with 630 nm reference. Cellular drug duration described as 48 hours. limitations: Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

experimental_model

Human NOZ and GBC-SD gallbladder cancer cells

exposure

Methods 2.2 describes 48-hour cellular drug treatments with refresh at 24 hours; exact concentration must be checked per figure and is not inferred from another assay.

limitations

Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

organism

Human

primary_locator

Figure 5B; Supplementary Figure S4B; Results 3.5.

primary_references

https://doi.org/10.1002/ptr.70245

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 96–102

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human NOZ and GBC-SD gallbladder cancer cells · source_derived_draft · unverified_draft

Solasonine did not change APT2 protein expression in the reported immunoblots. primary_references: https://doi.org/10.1002/ptr.70245 primary_locator: Figure 5B; Supplementary Figure S4B; Results 3.5. organism: Human experimental_model: Human NOZ and GBC-SD gallbladder cancer cells exposure: Methods 2.2 describes 48-hour cellular drug treatments with refresh at 24 hours; exact concentration must be checked per figure and is not inferred from another assay. limitations: Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

experimental_model

Human NOZ and GBC-SD gallbladder cancer cells

exposure

25 micromolar solasonine; main methods specify 48-hour cellular drug exposure.

limitations

Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

organism

Human

primary_locator

Figure 5D-E; Results 3.5.

primary_references

https://doi.org/10.1002/ptr.70245

APT2, STAT3 and GPX4: opposing branches and assay qualification · lines 114–120

Primary observations: 10.1038/s41467-025-56344-5 and 10.1002/ptr.70245; selected full-text review 2026-09-20. · supports · Human NOZ and GBC-SD gallbladder cancer cells · source_derived_draft · unverified_draft

Solasonine reduced APT2-STAT3 co-immunoprecipitation. primary_references: https://doi.org/10.1002/ptr.70245 primary_locator: Figure 5D-E; Results 3.5. organism: Human experimental_model: Human NOZ and GBC-SD gallbladder cancer cells exposure: 25 micromolar solasonine; main methods specify 48-hour cellular drug exposure. limitations: Human cancer-cell experiments; no clinical or dietary inference. Solasonine exposure is not equivalent to APT2 depletion, ML349, or sulforaphane. Mutant rescue does not by itself establish selectivity or normal baseline mutant function. Figure 5C calls this APT2 activity, but Methods 2.17 describes an anti-LYPLA2 ELISA with mass-concentration standards and an HRP/TMB readout from cell supernatants. No direct APT2 substrate-turnover assay is described there. The result is retained as an author-labelled activity proxy, not established catalytic inhibition. This methodological qualification does not invalidate the separately reported binding or cellular observations.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0

experimental_model

Human HeLa; tagged human APT2-C2S and ZDHHC6 constructs

exposure

24-hour plasmid expression, then 2-hour tritiated-palmitate labeling; normalized to ZDHHC6 protein; n=6.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. This mutant result supports a role for APT2 palmitoylation in this substrate assay; it is not a sulforaphane experiment.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 2E,F; Rapid APT2-mediated ZDHHC6 depalmitoylation

primary_references

https://doi.org/10.7554/eLife.27826

source_access

Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 85–93

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HeLa; tagged human APT2-C2S and ZDHHC6 constructs · source_derived_draft · unverified_draft

Palmitoylation-deficient APT2-C2S overexpression did not reproduce the significant reduction in ZDHHC6 palmitoylation caused by wild-type APT2. primary_references: https://doi.org/10.7554/eLife.27826 primary_locator: Figure 2E,F; Rapid APT2-mediated ZDHHC6 depalmitoylation source_access: Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0 experimental_model: Human HeLa; tagged human APT2-C2S and ZDHHC6 constructs organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 24-hour plasmid expression, then 2-hour tritiated-palmitate labeling; normalized to ZDHHC6 protein; n=6. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. This mutant result supports a role for APT2 palmitoylation in this substrate assay; it is not a sulforaphane experiment.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a

experimental_model

Human HEK293 expressing FLAG-tagged APT2-C56S

exposure

100 micromolar sulforaphane for 3 hours followed by 50 micromolar Alk14 for 5 hours; three separate experiments.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. C56S is a mutant-plus-exposure actor. Its baseline function must be assessed before using it as a selective pharmacological rescue.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 4C

primary_references

https://doi.org/10.1016/j.bbrc.2024.150244

source_access

Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 41–49

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HEK293 expressing FLAG-tagged APT2-C56S · source_derived_draft · unverified_draft

Sulforaphane did not significantly attenuate Alk14 labeling of APT2-C56S under the conditions that reduced labeling of wild-type APT2. primary_references: https://doi.org/10.1016/j.bbrc.2024.150244 primary_locator: Figure 4C source_access: Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a experimental_model: Human HEK293 expressing FLAG-tagged APT2-C56S organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 100 micromolar sulforaphane for 3 hours followed by 50 micromolar Alk14 for 5 hours; three separate experiments. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. C56S is a mutant-plus-exposure actor. Its baseline function must be assessed before using it as a selective pharmacological rescue.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0

experimental_model

Human HeLa; LYPLA2 siRNA and tagged ZDHHC6

exposure

72-hour siRNA protocol; 2-hour metabolic protein pulse followed by chase. Reported apparent half-lives, not raw-data refits.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. C328 mutation abolished sensitivity and MG132 rescued degradation in the reported experiments. Higher regulatory palmitoylation is not equivalent to sustained higher ZDHHC6 abundance.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 3A-E; ZDHHC6 palmitoylation controls degradation

primary_references

https://doi.org/10.7554/eLife.27826

source_access

Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 96–104

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HeLa; LYPLA2 siRNA and tagged ZDHHC6 · source_derived_draft · unverified_draft

LYPLA2 silencing accelerated ZDHHC6 decay in HeLa pulse-chase experiments, shortening the reported apparent half-life from about 16 hours to about 3 hours. primary_references: https://doi.org/10.7554/eLife.27826 primary_locator: Figure 3A-E; ZDHHC6 palmitoylation controls degradation source_access: Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0 experimental_model: Human HeLa; LYPLA2 siRNA and tagged ZDHHC6 organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 72-hour siRNA protocol; 2-hour metabolic protein pulse followed by chase. Reported apparent half-lives, not raw-data refits. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. C328 mutation abolished sensitivity and MG132 rescued degradation in the reported experiments. Higher regulatory palmitoylation is not equivalent to sustained higher ZDHHC6 abundance.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc

experimental_model

Human A375 and HT1080 cancer cells; lentiviral APT2 shRNA

exposure

RT-qPCR; three independent experiments; exposure timing unresolved.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The actor names the A375 arm; the parallel HT1080 observation is preserved in the context. No transcriptional effect is inferred.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 5f

primary_references

https://doi.org/10.1038/s41467-025-56344-5

source_access

Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 129–137

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human A375 and HT1080 cancer cells; lentiviral APT2 shRNA · source_derived_draft · unverified_draft

APT2 knockdown did not significantly change GPX4 mRNA in the reported A375 and HT1080 RT-qPCR experiments. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 5f source_access: Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc experimental_model: Human A375 and HT1080 cancer cells; lentiviral APT2 shRNA organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: RT-qPCR; three independent experiments; exposure timing unresolved. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The actor names the A375 arm; the parallel HT1080 observation is preserved in the context. No transcriptional effect is inferred.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc

experimental_model

Human A375 melanoma; lentiviral APT2 shRNA; exogenous GPX4

exposure

ABE assay with and without hydroxylamine; representative of three independent experiments; exact shRNA exposure duration unresolved.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The Results prose calls this deletion, but Figure 5d specifies shRNA; record the figure-defined knockdown, not a knockout.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 5d and Results: APT2 mediates the depalmitoylation of GPX4

primary_references

https://doi.org/10.1038/s41467-025-56344-5

source_access

Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 107–115

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human A375 melanoma; lentiviral APT2 shRNA; exogenous GPX4 · source_derived_draft · unverified_draft

APT2 shRNA increased GPX4 palmitoylation measured by acyl-biotin exchange in A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 5d and Results: APT2 mediates the depalmitoylation of GPX4 source_access: Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc experimental_model: Human A375 melanoma; lentiviral APT2 shRNA; exogenous GPX4 organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: ABE assay with and without hydroxylamine; representative of three independent experiments; exact shRNA exposure duration unresolved. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The Results prose calls this deletion, but Figure 5d specifies shRNA; record the figure-defined knockdown, not a knockout.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc

experimental_model

Human A375 melanoma; lentiviral APT2 shRNA

exposure

Cycloheximide chase; three independent experiments; exact CHX concentration and time series not resolved from reviewed legend.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 5i,j

primary_references

https://doi.org/10.1038/s41467-025-56344-5

source_access

Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 118–126

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human A375 melanoma; lentiviral APT2 shRNA · source_derived_draft · unverified_draft

APT2 knockdown increased GPX4 protein stability in A375 cycloheximide-chase experiments. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 5i,j source_access: Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc experimental_model: Human A375 melanoma; lentiviral APT2 shRNA organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: Cycloheximide chase; three independent experiments; exact CHX concentration and time series not resolved from reviewed legend. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements.

availability_state

machinery_impairment Imported condition classification; unverified.

evidence_cache

artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc

experimental_model

Human A375 melanoma; lentiviral APT2 shRNA

exposure

4 micromolar RSL3 for 6 hours; SYTOX Green staining; three independent experiments.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. This is a response to a GPX4 inhibitor in cancer cells. It does not establish an effect of sulforaphane or dietary selenium.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 6c,d

primary_references

https://doi.org/10.1038/s41467-025-56344-5

source_access

Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis.

trigger_kind

machinery_impairment Imported condition classification; unverified.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 140–148

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human A375 melanoma; lentiviral APT2 shRNA · source_derived_draft · unverified_draft

APT2 knockdown reduced RSL3-induced cell death measured with SYTOX Green in A375 cells. primary_references: https://doi.org/10.1038/s41467-025-56344-5 primary_locator: Figure 6c,d source_access: Selected primary Results and Figure 5/6 legends reviewed. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/gpx4-primary-passages.json; SHA256 384aaae29ec0e6fe9814782b3e5d885753751fa1b2bbd103e30e8ffa113df5dc experimental_model: Human A375 melanoma; lentiviral APT2 shRNA organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 4 micromolar RSL3 for 6 hours; SYTOX Green staining; three independent experiments. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. This is a response to a GPX4 inhibitor in cancer cells. It does not establish an effect of sulforaphane or dietary selenium.

evidence_cache

artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0

experimental_model

Human HeLa; tagged human APT2 and ZDHHC6 constructs

exposure

24-hour plasmid expression, then 2-hour tritiated-palmitate labeling; normalized to ZDHHC6 protein; n=6.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 2E,F; Rapid APT2-mediated ZDHHC6 depalmitoylation

primary_references

https://doi.org/10.7554/eLife.27826

source_access

Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 74–82

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HeLa; tagged human APT2 and ZDHHC6 constructs · source_derived_draft · unverified_draft

Wild-type APT2 overexpression reduced radiolabeled palmitate incorporation into ZDHHC6 in HeLa cells. primary_references: https://doi.org/10.7554/eLife.27826 primary_locator: Figure 2E,F; Rapid APT2-mediated ZDHHC6 depalmitoylation source_access: Selected primary Results, figure legends and methods via indexed publisher text. No supplement or raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/zdhhc6-primary-passages.json; SHA256 a02040bb8922a9170e095cdfb5e875a5e7f513b682c0ec69c19feacb3dc191b0 experimental_model: Human HeLa; tagged human APT2 and ZDHHC6 constructs organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 24-hour plasmid expression, then 2-hour tritiated-palmitate labeling; normalized to ZDHHC6 protein; n=6. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements.

evidence_cache

artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a

experimental_model

Human HEK293 expressing FLAG-tagged human APT2

exposure

100 micromolar sulforaphane for 3 hours; membrane and cytosol fractions with pan-cadherin and alpha-tubulin markers; three separate experiments.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The authors describe plasma-membrane localization, but fractionation is not an ER-specific or organelle-resolved measurement.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 4A; Results 3.4 and Methods 2.9

primary_references

https://doi.org/10.1016/j.bbrc.2024.150244

source_access

Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 30–38

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HEK293 expressing FLAG-tagged human APT2 · source_derived_draft · unverified_draft

Sulforaphane reduced the membrane-to-cytosol ratio of FLAG-tagged APT2 in HEK293 fractionation experiments. primary_references: https://doi.org/10.1016/j.bbrc.2024.150244 primary_locator: Figure 4A; Results 3.4 and Methods 2.9 source_access: Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a experimental_model: Human HEK293 expressing FLAG-tagged human APT2 organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 100 micromolar sulforaphane for 3 hours; membrane and cytosol fractions with pan-cadherin and alpha-tubulin markers; three separate experiments. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The authors describe plasma-membrane localization, but fractionation is not an ER-specific or organelle-resolved measurement.

evidence_cache

artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a

experimental_model

Human HEK293 expressing FLAG-tagged human APT2

exposure

100 micromolar sulforaphane for 3 hours followed by 50 micromolar Alk14 for 5 hours; hydroxylamine control; three separate experiments.

limitations

These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The authors could not distinguish reduced palmitoylation from enhanced depalmitoylation. This assay does not measure GPX4 or ZDHHC6.

organism

Human experimental cells; construct species unresolved where stated for discussion-only nulls

primary_locator

Figure 4B; Methods 2.8

primary_references

https://doi.org/10.1016/j.bbrc.2024.150244

source_access

Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis.

APT2: sulforaphane engagement, GPX4 stability and ZDHHC6 turnover · lines 19–27

Targeted primary-literature curation from recursive ZDHHC6 exploration, 2026-09-20. · supports · Human HEK293 expressing FLAG-tagged human APT2 · source_derived_draft · unverified_draft

Sulforaphane reduced Alk14 labeling of immunoprecipitated APT2 in transfected HEK293 cells. primary_references: https://doi.org/10.1016/j.bbrc.2024.150244 primary_locator: Figure 4B; Methods 2.8 source_access: Full six-page primary PDF, methods and discussion read; Figure 4 visually inspected. No raw-data reanalysis. evidence_cache: artifacts/discovery-research/round6-sources/sulforaphane-apt2.pdf; SHA256 31e2993f88fe05eec199b9013173b29379b3346b9696ac766771173ddd0f181a experimental_model: Human HEK293 expressing FLAG-tagged human APT2 organism: Human experimental cells; construct species unresolved where stated for discussion-only nulls exposure: 100 micromolar sulforaphane for 3 hours followed by 50 micromolar Alk14 for 5 hours; hydroxylamine control; three separate experiments. limitations: These are separate experimental observations, not a demonstrated sulforaphane-GPX4-ZDHHC6 pathway. Sulforaphane-driven APT2 relocalization is not equivalent to genetic depletion or general catalytic inhibition. No dietary, clinical or selenium-repletion effect is inferred. Protein stability, palmitoylation and substrate output remain different measurements. The authors could not distinguish reduced palmitoylation from enhanced depalmitoylation. This assay does not measure GPX4 or ZDHHC6.