Fig. 2

NAC induces apoptosis after sublethal heat stress. HepG2 and HuH7 cells that were untreated or subjected to sublethal heat stress were cultured for 12 h in medium with or without NAC (5 mM). A and B Mitochondrial membrane potential was assessed by TMRE staining and analyzed by flow cytometry at 12 h after sublethal heat stress. C and D Cell death was quantified using Annexin V-FITC/PI staining and flow cytometry at 24 h after sublethal heat stress. Summary graph representing the percentage of single PI + cells (Q1 areas) (E) and Annexin V + cells (Q2 and Q3 areas) (F) in each group. G Western blot of GAPDH, BAX, BCL-2, and Cleaved-caspase-3 expression. H mRNA expression of RIPK1 and RIPK3.HepG2 and HuH7 cells incubated with NAC(5 mM) at 12 h after sublethal heat stress. (I)Cell death was quantified using Annexin V-FITC/PI staining and flow cytometry at 24 h after sublethal heat stress. Values are the mean ± SE.**p < 0.01; ***p < 0.001; ns no significance