Research Paper Volume 12, Issue 23 pp 24318—24332

Figure 5. Autophagy inhibition attenuates ISO-induced cell death. HepG2 and Huh7cells were pre-treated with 3-MA (5 mM) or vehicle for 2 h or transfected with si-Atg5 or si-NC for 6 h, and then treated with ISO (400 μM) for 48 h. (A, D) CellTiter-Blue viability assay results. Data are expressed as percentage relative to untreated control cells. (B, E) Western blotting analysis 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.