Figure 3

Non-phosphorylatable Puma mutant S10A induces apoptosis at a greater rate than WT-Puma in HeLa cells. (a and b) HeLa cells were co-transfected with the indicated Puma construct and a construct encoding farnesylated GFP. At 16 h after transfection, cells were fixed with 4% PFA for 20 min, DNA was stained with Hoechst 33342 and then coverslips mounted for analysis by fluorescence microscopy (a). Cell death was quantified by examining the nuclear morphology of GFP-positive cells and calculating the percentage displaying condensed or fragmented apoptotic nuclei (white arrowheads) (b). ^**P<0.01