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Figure 2 Detection of apoptosis by annexin V–FITC binding. Cardiac myocytes were treated with serum-free DMEM alone (a), 10^–6 mol/l of doxorubicin (b), 10^–6 mol/l of doxorubicin in the presence of 10^–6 mol/l of amlodipine (c), or 10^–6 mol/l of nifedipine (d) for 14 h. (A) The cells were then stained with annexin V–FITC and visualized by fluorescence microscopy, as described in the Methods section. Photomicrographs are representative of at least three experiments for each experiment (magnification x 400). (B and C) Quantification of apoptosis in myocytes was determined by flow cytometric analysis. Cells were harvested, stained with annexin V–FITC, and analyzed as described in Methods. The percentage of apoptotic myocytes was determined by calculating the ratio of annexin V–FITC binding cells (M1) to total cells (n = 6). *p < 0.001 vs. control. p < 0.001 vs. doxorubicin.