Figure 3

10058-F4 induced apoptosis in ovarian cancer cells. SKOV3 and Hey cells were treated with 10058-F4 at the indicated doses for 24 hours and then analyzed for AnnexinV and PI staining by Cellometer (A and B). The measurement of early and late apoptosis was shown in C (Hey) and D (SKOV3) at two independent experiments. Hey and SKOV3 cells treated with 10058-F4 for 16 hours were analyzed by Western blotting using PARP, capspase3, caspase7, BCL-2, MCL-1 and β-actin antibodies (E). The BCl-2 and MaCL-1 were analyzed by Western blotting after 16 hours treatment with 10058-F4. 10058-F4 increased cleaved PARP, caspase3 and caspase7 protein expression (F and G) and decreased BCL-2 and MCL-1 protein expression (H). Caspase-3 activity (I) and cell proliferation (J) were determined by ELISA and MTT assay after the both cells were treated with Z-VAD-FMK, 10058-F4 or combination for 24 or 48 hours. *p < 0.05 and **p < 0.01, two-sided student t test.