fig3

Figure 3. Withaferin A induced apoptosis in floating (F) cells. (a) Using cytofluorometric technique and Annexin V as an apoptotic indicator, we demonstrated that withaferin A promoted apoptosis in parental (P), F and adherent (A) cells in a dose-dependent manner; (b) quantitative representation of withaferin A-induced apoptosis. Withaferin A appeared to trigger apoptosis to a higher extent in F cells than in P cells (P < 0.01) or in A cells (P < 0.01); (c) immunoblots of total cell lysates obtained from withaferin A-treated F cells showed an increased expression of pro-apoptotic molecules such as caspase-3, -8, -9, and poly ADP-ribose polymerase at higher concentrations of withaferin A (5-10 μmol/L), except Fas receptor. On the other hand, anti-apoptotic molecules, survivin, and Bcl-2 were clearly down-regulated when the two higher concentrations of withaferin A were used