Figure 6
4bHWE arrested the cell cycle at the G2/M phase and induced apoptosis. Huh-7 cells were treated with 4bHWE at the indicated concentration for 24 h and then stained with PI. (a) DNA content was analyzed using flow cytometry. (b) Histogram of the cell cycle distribution. (c) Histogram of the apoptotic sub-G1 population. (d) Huh-7 cells were treated with the indicated concentration of 4bHWE for 24 h and then harvested. Equal amounts of whole cell lysates (20 μg) were separated using SDS-PAGE and immunoblotted with various antibodies as indicated. Tubulin is shown as an internal standard. The fold-change values are presented below each band. (e) Effect of a pan-caspase inhibitor on 4bHWE-induced apoptosis in Huh-7 cells. Cells were pretreated with 20 μM pan-caspase inhibitor for 1 h, and then treated with 10 μM 4bHWE for 24 h. Cell viability was determined using MTT assay. Data are the mean of three independent experiments. *p < 0.05 compared with the control. Full-length blots are presented in Supplementary Fig. S4.
