Figure 4.

WIN induces apoptosis of U251 cells. (a) WIN time-dependently induces apoptosis of U251 cells. U251 cells seeded in 6-well plate were treated with 20 μM WIN for 0-12 h and loaded with Annexin V probe for 20 min in darkness. Then, cells were washed and the cell apoptosis were imaged by a fluorescence microscope (magnification, ×200). (b) Quantitative analysis of cell apoptosis. (c) WIN induced caspase-3 activation and Cyto-C release. Protein expression was detected by western blotting assay. All experiments were repeated three times. Bar with “^∗” or “^∗∗” indicates the significant difference at P < 0.05 or P < 0.01 level, respectively.