Figure 2.

Danu treatment arrested Hep3B cells in G₂/M phase via regulating the expression of key cell cycle regulators. 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 p-CDC25C (Ser216), p-CDC2/CDK1 (Tyr15), CDC2/CDK1, p-cyclin B1 (Ser133), and cyclin B1 (B). Bar graphs show the percentage of Hep3B cells in G₁, S, and G₂/M phases 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.