Figure 3.

Danu treatment induced apoptosis via mitochondria-dependent pathway. Hep3B cells were treated with Danu at 0.01, 0.1, and 0.5 µM for 24 hr, and then subject to flow cytometric analysis. The protein samples were subject to Western blotting assay. Representative flow cytometric plots of cell cycle distribution (A) and blots of cytochrome c, Bax, Bcl-xl, Bcl-2, cleaved caspase 9, cleaved caspase 3, PUMA, and cleaved PARP (B). Bar graphs show the percentage of autophagic Hep3B cells and the relative level of the above proteins. β-Actin was used as the internal control. Data are the mean ± SD of three independent experiments. *P<0.05, **P<0.001 and ***P<0.001 by one-way ANOVA.