Evaluation of apoptosis by Annexin-V method. Apoptosis was assessed after incubating the cells with Gli or vehicle by 72 h. For positive control cells were treated with H2O2 (5 mM) for 30 minutes. Fluorescence was evaluated immediately after Annexin-V staining by flow cytometry. Gli (25 μM) did not induce an increment in apoptosis of MDA-MB-231 cells. Positive Annexin-V cells are shown in both right quadrants.