Figure 6. ISO-triggered autophagy in HCC cells is promoted by rapamycin and not prevented by Z-VAD-FMK. HepG2 and Huh7cells were treated with 400 μM ISO for 48 h in the presence or absence of 100 nM rapamycin (RAPA) or 100 μM z-VAD-FMK (zVAD). (A, D) CellTiter-Blue viability assay results. (B, E) Immunoblot analyses of LC3, p62, p-mTOR, mTOR, p-AMPK, AMPK, p-p70S6K, p70S6K, Bax, Bcl-2, cleaved caspase-3, and cleaved PARP expression. (C, F) Densitometric analysis of protein expression data. Data are expressed as fold change relative to values from untreated control cells after normalization against GAPDH. Values are expressed as mean ± SD; *P < 0.05; **P < 0.01; ***p < 0.001.