Fig. 4: Sensitivity to APAP-induced toxicity depends on mitochondrial energy production.
From: Non-canonical BIM-regulated energy metabolism determines drug-induced liver necrosis
A Seahorse Basal ATP Rate Assay of untreated primary murine hepatocytes (PMH) and IHH with glyco/mitoATP ratio above, n = 6–8. B CellTiter-Glo Assay of PMH and IHH cells treated for 6 h, n = 4–9. C Working hypothesis: Glucose withdrawal shifts cell lines to oxidative phosphorylation (OxPhos) dependency and elongated mitochondria (green), while MFN1/2 knockout reverses the phenotype. D Representative confocal images of MitoTracker Green-stained untreated IHH and HepG2 cultured in different glucose concentrations. Scale bar 20 µm. E Seahorse Mito Stress Test Assay of IHH (left) or Basal ATP Rate Assay of HepG2 (mid+right) cultured in normal (NG) or no glucose (noG). Basal respiration and maximal ECAR surrogate for mitochondrial and glycolytic activity. Statistical significance tested by comparing mitoATP and glycoATP independently. n = 3 (all). F FACS analysis of AnnexinV-FITC + IHH and HepG2 treated with 20 mM APAP for 24 h and cultured in NG or noG. n = 3–6 (left), n = 4 (mid), n = 4 (right). Data points and/or bar graphs are mean +/− SD with n as independent biological replicates. Statistical significance was tested using unpaired Student’s t test (E) or Two-way ANOVA with Sidak’s multiple comparison test (B, F).
