The indicated cell lines were transiently transfected with DNA to express VBimL, and the indicated VBimL mutant proteins. Cells expressing Venus fusion proteins were stained with the nuclear dye Draq5 and rhodamine-labeled Annexin V, and apoptosis was assessed by confocal microscopy as in Figure 1. The y-axis indicates Annexin V Positivity (%), which was calculated based on the total number of Venus expressing cells that also score positive for Annexin V rhodamine fluorescence. A minimum of 400 cells were imaged for each condition. Individual points (open circles) represent the average for each replicate, while the bar heights, relative to the y-axis, represent the average for all three replicates. A one-way ANOVA was used within each cell line followed by a Tukey’s multiple comparisons test to compare the means of each transfection group. *p-values<0.05, **p-values<0.01, ns, non-significant p-values (>0.05).
Figure 10—source data 1. Source data demonstrating that mutants of BimL that cannot activate Bax do not kill H.EK293 cells.