Figure 2.

Withagulatin A induces cell cycle arrest. (A) Withagulatin A arrests cells at S phase. LX-2 cells were incubated with Withagulatin A (0, 1, and 2 μmol/L) for 24 h. Cells were fixed, stained with propidium iodide, and analyzed by flow cytometry as described in Materials and methods. The experiment was repeated three times and representative data are shown. (B) Withagulatin A affects cell cycle regulatory proteins. LX-2 cells were incubated with Withagulatin A (2 μmol/L) for the indicated time points (0–24 h). Cells were harvested and cell lysates were analyzed by Western blotting for cyclin D1, p-CDK2, CDK2, and cyclin B1 levels. GAPDH was used as a loading control. Cells treated with DMSO were used as controls for each time point. Results shown are representative of three independent experiments. (C) Bands were quantified and data are expressed as fold of control. ^bP<0.05, ^cP<0.01 vs control cells.