Fig. 5

IC2 induces apoptosis of MCF-7 cells. MCF-7 cells were treated with three concentrations of IC2 for 24 h. A Representative images of DAPI staining at ×200 magnification in MCF-7 cells. B MCF-7 cells were double-stained with Annexin V and PI followed by flow cytometry analysis. The gate setting distinguished among living (bottom left), necrotic (top left), early apoptotic (bottom right) and late apoptotic (top right) cells. C Percentage of apoptotic cells compared to the control from B. D The expression of PARP, Bcl-2 and Bax was determined by Western blot. E, F Quantification of the protein expression levels of cleaved PARP/PARP and Bcl-2/Bax. Data are presented as the mean ± SEM of three independent experiments (n = 3). *P < 0.05, **P < 0.01 and ***P < 0.001 vs. control