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. 2010 Jul 26;285(40):30634–30643. doi: 10.1074/jbc.M110.116681

FIGURE 4.

FIGURE 4.

Effect of PMCA2 overexpression on cells viability and caspase-3 cleavage. a, cell viability. Nontransfected BRIN BD-11 cells (Ctrl) or different clones of BRIN cells transfected with PMCA2, clone 2 (Cl 2), or clone 5 (Cl 5) and/or aequorin targeted to the cytosol (Cyt-Aeq) or the mitochondria (mutated aequorin (Mit-Aeq)) were untreated (black bars) or treated for 24 h with CPA (white bars), thapsigargin (striped bars), or the solvent DMSO (dotted bars). Apoptosis levels were evaluated by observation under a microscope after Hoechst 33342-propidium iodide staining. The data are expressed as the percentage of apoptotic cells over the total number of cells counted ± S.E. Results are means of three to five independent experiments. *, p < 0.05 and **, p < 0.01 versus respective nontransfected control. b, caspase-3 cleavage. Western blot analyses of nontransfected (Ctrl) and PMCA2-transfected cells, clone 2 (Cl 2) or clone 5 (Cl 5) using an antibody directed against the cleaved caspase-3 fragment. Upper panel, representative blot of caspase-3 and β-actin expression. Lower panel, quantitative assessment of cleaved caspase-3 levels normalized to the β-actin levels. Results are means of five independent experiments. *, p < 0.05 versus respective nontreated control. #, p < 0.05 versus respective nontransfected condition.