Figure 2

Effects of co-treatment with I3C and genistein on apoptosis in HT-29 cells. A, The sub-G1 population was assessed by flow cytometry as described in Materials and Methods after exposure to the indicated agents for 24 or 48 h. Columns, mean (n = 3); bars, SD. *, P < 0.05, significantly different compared with the DMSO-treated control. B, HT-29 cells were exposed to DMSO or a combination of I3C (300 μmol/L) and genistein (40 μmol/L) for 48 h or pretreated with z-VAD-fmk (20 or 100 μmol/L) for 2 h followed by exposure to the combination for 48 h. The sub-G1 population was quantified by flow cytometry as described in Materials and Methods. Columns, mean (n = 3); bars, SD. *, P < 0.05, significantly different compared with the combined treatment. C, Cells were exposed to DMSO (control), a combination of I3C (300 μmol/L) and genistein (40 μmol/L), or doxorubicin (1 μmol/L) for 48 h, stained with DAPI, and observed under a fluorescence microscope as described in Materials and Methods. Doxorubicin was used as a positive control to induce apoptosis. Bars, 20 μm. D, HT-29 cells were exposed to the indicated agents for 48 h. The expression of PARP, caspase-8, caspase-9 and cleaved caspase-3 proteins was analyzed by western blotting. β-actin was used as a loading control. -, treated with DMSO. GEN, genistein.