Fig. 3

Epoxyazadiradione induces apoptosis through disruption of mitochondrial homeostasis: a MDA-MB-231 cells were treated with increasing concentration of epoxyazadiradione for 24 h. Cells were stained with DHE at 37 °C for 20 min and fluorescence intensity was analyzed by flow cytometry and represented as histogram. The percentage of DHE staining as compared to unstained cells was quantified and represented graphically. b MDA-MB-231 cells were treated with epoxyazadiradione for 24 h at indicated concentrations, stained with anti-AIF antibodies and analyzed by confocal microscopy. Nuclei were stained with DAPI. c MDA-MB-231 (upper panel) and MCF-7 (lower panel) cells were independently treated with epoxyazadiradione with different doses (0–150 μM) and JC-1 staining was performed at 37 °C for 20 min. JC-1 aggregates (red fluorescence) and JC-1 monomers (green fluorescence) in stained cells were analyzed by flow cytometry. The ratio of JC-1 aggregates and JC-1 monomers was analyzed statistically and represented graphically from three independent experiments. Values are represented in mean ± SEM. *, p < 0.007; #, p < 0.0008 by Student’s t test