Figure 4

Elevated autophagy level by rapamycin reduces apoptosis of PA-induced in hepatocytes. (A) Cells were treated with either control or PA (500 μM) for 24h, Rapa (250 nM) was added to pretreatment for 8 hours, then western blotting analysis tested LC3 and cleaved caspase3 expressions levels. (B) The quantization of relative LC3-II/GAPDH from the treatment groups were calculated by normalizing to the control groups. (C) Relative cleaved-caspase3/GAPDH was calculated in the same way. Data were presented as mean ± SEM from three independent experiments (*p < 0.05; **p < 0.01). (D) Cells were treated with control, PA, Rapa (250 nM) and PA with the addition of Rapa pretreatment for 24 hours, then cell viability was detected by CCK-8 analysis. Data were mean ± SEM from the three times independent experiments (*p < 0.05; **p < 0.01). (E) Cells were stained with AnnexinV-FITC/PI after treatment as above, and were measured by FCM. The results were presented as mean ± SEM values at least three independent experiments (*p < 0.05; **p < 0.01). (F) HL-7702 cells were treated with various strategies, and then CCK-8 detected cell proliferation. (G) The result of cell proliferation in HepG2 cells was shown. Date were presented as the mean ± SEM values for three independent experiments (*p < 0.05; **p < 0.01).