Fig. 7.

BNIP3 expression and caspase-independent cell death. (A, B) Cells were transfected with siRNA control or siRNA specific for BNIP3 and 24 h post-transfection cells were treated with 400 μM KCN for 24 h. BNIP3 expression as determined by Western blotting and apoptotic cell death by Hoechst 33258 staining. (C) Cells were transfected with EV control or BNIP3 cDNA for 24 h, followed by 400 μM KCN treatment for 24 h and then caspase-3 activity was determined. H₂O₂ (100 μM) treatment group was used as positive control. (D) Cells were transfected as in (C), followed by exposure to 400 μM KCN for 24 h in the presence or absence of zVAD-fmk (25 μM). Apoptosis was then determined. Data represent mean ± SEM of three to six separate determinations. *Significantly different from control group; #Signficantly different from KCN+RNAi control group. P<0.05.