Bax clustering under apoptotic stress.

<p>A) Bax clustering- MCF-7 cells stably expressing GFP-Bax were incubated 6 hours at 37°C with the different conditions and nuclei were stained with Hoechst (100 ng/mL). Here is shown a representative example of basal levels of Bax activation (BSS) and an example of Bax activation under camptothecin (2 µM). B) Active Bax translocates to the mitochondria- MCF-7 cells stably expressing GFP-Bax were transiently transfected with mito-mCherry and incubated 6 hours (37°C) with camptothecin (2 µM) (Hoechst for nuclei). The 3D rendering (ImageJ) image shows GFP-Bax (green) translocated to mitochondria (in red). C) Bax clustering- Representative microscope region for each pro-apoptotic condition is shown. D) Bax levels- Cells with GFP-Bax clusters were scored as “positive” for Bax activation. D) Cells “positive” for activated Bax were scored and plotted as shown. Values are presented as mean percentage ± s.e.m. (N = 5, approx. 500 cells/condition; *, P≤0.05, * *, P≤0.01, t-test). Images were acquired with a DVRT scope and a 40× Objective.