Figure 2

PA stimulates autophagy activation in hepatocytes. (A) HL-7702 cells were treated with control or PA for 24 hours, and plasmids of GFP-LC3 were transfected into the cells. Cells were observed under fluorescence microscope (bar: 20μm). Quantization was obtained by calculating the ratio of cells with GFP-LC3 dots in one visual field and experiments were repeated three times (*p < 0.05; **p < 0.01). (B) LC3 and P62 protein levels were detected by western blotting analysis after treatment of control or PA for 8 hours. (C) Cells were treated with control or PA for 8 hours before being processed, then electron microscope was performed at 11,500× and 29,500× magnification. The black arrows show membrane-bound vacuoles characteristic of autophagosomes. The number of autophagosomes per cell was quantitated. Date were presented as the mean ± SEM of three independent experiments (*p < 0.05; **p < 0.01).