Fig. 4.

Epoxyazadiradione augments apoptosis in breast cancer cells through Caspase-dependent and ROS independent pathway: a MDA-MB-231 cells were treated with epoxyazadiradione (0–150 μM) for 24 h and the expression of Bcl2 family proteins including Bax, Bad and Bcl2 and cleaved caspase 9, 3 and PARP were analyzed by immunoblot. Actin was reprobed with each respective original blot and used as loading control. b The expression of Bcl2 family protein including Bax and Bcl2 and PARP were analyzed by western blot in MCF-7 cells upon treatment with epoxyazadiradione at indicated doses. c The ratios of Bcl2 with Bad or Bax were represented by densitometric quantification of western blot from three independent experiments in MDA-MB-231 cells. d The ratio of Bcl2 and Bax was represented by densitometric quantification of western blot from three independent experiments and represented graphically in MCF-7 cells. e MDA-MB-231 cells were either independently pretreated with caspase 9 inhibitor I (20 and 40 μM) or N-acetyl cysteine (NAC, 2.5 and 5 mM) or catalase (200 and 400 U/ml) for 1 h followed by treatment with epoxyazadiradione (150 μM) for another 24 h. The cell viability was analyzed using MTT assay. Values are represented in mean ± SEM of three independent experiments; *, p < 0.00004