Berberine induces apoptotic and necrotic death of HepG2 cells . HepG2 cells were either left untreated or treated with described concentration of berberine (10, 50, 100 and 200 μM), cells were further cultured in DMEM (no serum) for 24 hours, expressions of Beclin-1, LC3B-II, Erk1/2 and β-actin were tested by western blots (A). The number of LC3-GFP puncta positive cells (autophagic cells) was counted (B).Cell viability of HepG2 cells with indicated treatment for 48 hours was tested by MTT assay (C). HepG2 cells transfected with scramble control siRNA, Beclin-1 siRNA or LC3B siRNA (100 nM each, for 48 hours) were either left untreated, or stimulated with berberine (100 μM) plus z-VAD-fmk (50 μM) (Berberine + ZVAD), cells were further cultured for 48 hours, expression of Beclin-1, LC3B and β-actin was tested by western blots (D), cell viability was also tested (E). Experiments in this figure were repeated three times, and similar results were obtained. Data were expressed as mean ± SD. #p < 0.05 vs. Ctrl group (B). *p < 0.05 (C and E).