Figure 3

Effect of ES, ES-BAX, ES-BAK, and ES-BAX-ES on endothelial cells. (a) C-PAE cells were incubated in the presence or absence of 20 μg/ml of either protein for 16 h. The cells were trypsinized and stained with propidium iodide. Fluorescence of individual nuclei was measured by flow cytometry. Each sample was analyzed in triplicate. Error bars: mean±S.E.M. The statistical significance of differences between the groups was assessed with one-way ANOVA with Bonferroni post-test, ***P<0.001. (b) Analysis of caspase-3 activation in C-PAE cells untreated or treated for 24 h with 20 μg/ml of each protein. β-Actin was used as a loading control. (c) C-PAE cells were incubated in the absence or presence of 3.12–50 μg/ml of either protein for 72 h, and relative cell viability was determined by the MTS assay. The mean absorbance of the control group at 495 nm was set as 100% viability. Each sample was analyzed in triplicate. (d) C-PAE cells were incubated in the absence or presence of 25 μg/ml of either protein for 72 h, and relative cell viability was determined by the MTS assay. (e) C-PAE cells were incubated in the presence or absence of 25 μg/ml of either protein for 45 min to 72 h, and relative cell viability was determined by the MTS assay. Error bars: mean±S.E.M. The statistical significance of differences between the groups was assessed by one-way ANOVA with Bonferroni post-test, **P<0.01; ***P<0.001