Figure 3 | Scientific Reports

Figure 3

From: Bafilomycin A1 induces caspase-independent cell death in hepatocellular carcinoma cells via targeting of autophagy and MAPK pathways

Figure 3

BafA1 triggers caspase-independent cell death.

BEL7402 and HepG2 cells were treated with vehicle control, 5 nM BafA1 or using 2 μM Doxorubicin (Dox) as a positive control. (A) After 48 h, cells were stained using Annexin V/PI double-staining and analyzed by FACS. (B) At 24, 48 and 72 h, Caspase-3, PARP and p53 expression was examined by IB, using β-actin as a loading control. (C) Cells were treated with BafA1 in the presence or absence of the caspase inhibitor, Z-VAD-FMK (50 μM) for 48 h double stained with Annexin V/PI and analyzed by FACS. (D,E) BEL7402 and HepG2 cells were treated for 24, 48 and 72 h using BafA1 and expression of Bcl2, Bcl-XL, Puma, Bid, Bax and Bim was examined by IB. (F) BEL7402 and HepG2 cells were transfected with two siPuma or siControl oligonucleotides for 48 h, treated with BafA1 for 24 h and cell viability was measured using the MTT assay. Data are presented as mean ± SEM from three independent experiments, (BafA1 means Bafilomycin A1).

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