Berberine induces apoptotic and necrotic death of HepG2 cells . HepG2 cells were either left untreated or treated with described concentration of berberine, cells were further cultured in DMEM for 48 hours, the cell viability was tested by “MTT” assay (A), the percentage of trypan blue dye positive cells was recorded (B); HepG2 cell proliferation was analyzed by BrdU incorporation assay (C). HepG2 cells treated with or without berberine were cultured in DMEM for 24 hours, apoptotic and necrotic cell death was tested by Annexin V FACS assay (D and E), expressions of cleaved-caspase 3, Bcl-2 and β-actin were tested by western blots (F). HepG2 cells were pre-treated with z-VAD-fmk (50 μM) for 1 hour, followed by berberine (50 and 100 μM) stimulation, cells were further cultured for 48 hours before cell viability was tested (G). 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 (A and B). #p < 0.05 vs. berberine-treated group (C).