Figure 3.

Withagulatin A induces cell apoptosis. (A) Induction of an apoptotic morphology by Withagulatin A. LX-2 cells were treated with Withagulatin A (0, 1, and 2 μmol/L) for 24 h. Cells were fixed, stained with Hoechst 33342, and visualized with a fluorescence microscope (scale bar=0 μm). Nuclear condensation and fragmentation are indicated by arrows. Representative views from five fields for each slide are shown and the treatments were performed in triplicate. (B) Withagulatin A affects apoptotic protein levels. LX-2 cells were treated with Withagulatin A (0, 1, and 2 μmol/L) for 24 h. Cells were harvested and lysates were analyzed by Western blot analysis to determine the levels of Bcl-2, Bax, cleaved-caspase3 (c-caspase3), and caspase3. GAPDH was used as a loading control. 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.