Research Paper Volume 12, Issue 23 pp 24318—24332

Figure 3. ISO induces autophagy in HCC cells. (A) Immunofluorescent detection of LC3 in HepG2 and Huh7 cells treated with ISO for 48 h. Nuclei were stained with DAPI. Scale bar: 100 μm. (B) The number of LC3 dots per cell were quantified using ImageJ. (C) Western blotting detection of autophagy-related proteins in HepG2 and Huh7 cells treated with ISO for 48 h. (D) Densitometric analysis of the data shown in (C). (E, F) Western blot analysis of LC3, p62, Bax, Bcl-2, cleaved caspase-3, and cleaved PARP expression in HepG2 and Huh7 cells treated with 400 μM ISO for 48, alone or after 2-h pre-treatment with 100 nM Bafilomycin A1. GAPDH was used as loading control. Values represent mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001.