{"id":"7e78ae55-4355-53ab-bcd1-7c48f08e9955","stable_key":"8c946991-97f0-55ee-8df0-00e67621962f:nad-plus-free-pools","predicate":"measured_in","statement":"A compartment-targeted biosensor estimated free NAD+ in HEK293T cells at 106 micromolar in cytoplasm, 109 micromolar in nucleus and 230 micromolar in mitochondria.","claim_class":"observational","status":"source_derived_draft","evidence_grade":"ungraded","direction":"context_dependent","is_public":true,"mechanism_event_id":"f43a5a73-23b2-5bdd-8b7d-83684c3cab51","mechanism_event_label":"Different compartments contain different amounts of freely available NAD+.","subject":{"id":"283ed24b-06a1-50aa-9281-df3bac6ce37e","slug":"nad-plus","display_name":"NAD+","entity_type_key":"small_molecule"},"object":{"id":"e88b4681-2e0b-5146-afc0-3983302d16c1","slug":"human-hek293t-free-nad-pools","display_name":"Free NAD+ pools in human HEK293T compartments","entity_type_key":"cellular_process"},"evidence_count":1,"mechanism_event":{"id":"f43a5a73-23b2-5bdd-8b7d-83684c3cab51","stable_key":"8c946991-97f0-55ee-8df0-00e67621962f:nad-plus-free-pools-event","event_type":"observed_relationship","label":"Different compartments contain different amounts of freely available NAD+.","description":"A compartment-targeted biosensor estimated free NAD+ in HEK293T cells at 106 micromolar in cytoplasm, 109 micromolar in nucleus and 230 micromolar in mitochondria.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"283ed24b-06a1-50aa-9281-df3bac6ce37e","slug":"nad-plus","display_name":"NAD+","entity_type_key":"small_molecule"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"e88b4681-2e0b-5146-afc0-3983302d16c1","slug":"human-hek293t-free-nad-pools","display_name":"Free NAD+ pools in human HEK293T compartments","entity_type_key":"cellular_process"},"role":"target","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""}]},"contexts":[{"dimension":"evidence_access","value_text":"Primary full text","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"Genetically encoded biosensor with permeabilization calibration in human HEK293T cells.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"These are cell-line estimates with calibration assumptions, not normal human blood ranges or dietary deficiency thresholds.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"NAD+ collection; molecular form, preparation, species, exposure and manipulation remain explicit.","comparator":null,"unit":null,"notes":"","entity":{"slug":"nad-plus","display_name":"NAD+","entity_type_key":"small_molecule"}},{"dimension":"plain_language","value_text":"Different compartments contain different amounts of freely available NAD+.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"Biosensor reveals multiple sources for mitochondrial NAD⁺. · 2016 · https://pubmed.ncbi.nlm.nih.gov/27313049/ · DOI 10.1126/science.aad5168","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"8ec8c53f-05e1-5573-ba38-fd7c9f019ce0","evidence_kind":"source_excerpt","locator":"Lines 12-18","start_line":12,"end_line":18,"excerpt":"## nad-plus-free-pools\nDifferent compartments contain different amounts of freely available NAD+.\nA compartment-targeted biosensor estimated free NAD+ in HEK293T cells at 106 micromolar in cytoplasm, 109 micromolar in nucleus and 230 micromolar in mitochondria.\nModel: Genetically encoded biosensor with permeabilization calibration in human HEK293T cells.\nLimitations: These are cell-line estimates with calibration assumptions, not normal human blood ranges or dietary deficiency thresholds.\nEvidence access: Primary full text\nBiosensor reveals multiple sources for mitochondrial NAD⁺. · 2016 · https://pubmed.ncbi.nlm.nih.gov/27313049/ · DOI 10.1126/science.aad5168","model_system":"Genetically encoded biosensor with permeabilization calibration in human HEK293T cells.","directness":"reported_statement","verification_status":"source_derived_draft","notes":"Original curation paraphrase; evidence access and experimental limitations specified.","relationship":"supports","weight":1.0,"link_notes":"","source":{"id":"f59aa302-d463-5462-876f-e21e64861eac","stable_key":"import-8c946991-97f0-55ee-8df0-00e67621962f","title":"NAD+: compartmental supply, consumption and cross-nutrient mechanisms (2026-09-19)","document_type":"imported_text","citation_label":"AI-assisted research curation; primary references, access levels and experimental limitations individually identified. 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