Figure 4

Detection of apoptosis induced by M22 by flow cytometry and confocal microscopy. (A) Evaluation of apoptosis by Annexin V/PI dual staining assay. Numbers of Annexin V-and PI-positive cells were determined using flow cytometry. Quadrant 1: necrotic cells; Quadrant 2: late-apoptotic cells; Quadrant 3: viable cells; Quadrant 4: early apoptotic cells. (B) The percent of apoptotic cells are represented by bar diagram. (C) Morphologic changes of A549 cells treated by M22 for 48 h were fixed and stained with Hoechst 33258. M22 treated cells show apoptotic, condensed and fragmented fluorescent nuclei. Bars, 10 μm.