Figure 2. ISO induces apoptosis in HepG2 and Huh7 cells. (A) Apoptosis detection by Annexin V/PI staining and flow cytometry in HepG2 and Huh7 cells treated with ISO for 48 h. Criteria were set to distinguish between viable (bottom left), early apoptotic (bottom right), late apoptotic (top right), and necrotic (top left) cells. (B) Statistical analysis of apoptosis rate. (C) Western blotting analysis of apoptosis-related proteins. GAPDH was used as loading control. (D) Densitometric analysis of the data shown in (C). Values represent mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001.