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. 2010 Oct 4;285(50):39108–39116. doi: 10.1074/jbc.M110.151753

FIGURE 5.

FIGURE 5.

MST1-induced histone H2AX phosphorylation is dependent on its cleavage in HeLa cells. A, HeLa cells were transfected with the pCE-MST1-FL or pCE-MST1-NT plasmid. The caspase-3 inhibitor Z-DEVD-fmk (20 μm) was added to the medium immediately after transfection. At 36 h after transfection, cells were harvested, and lysates and histone proteins were prepared. The expression level of MST1-FL or MST1-NT was detected with anti-Myc-Tag, and the level of γ-H2AX expression was detected in the acid-extracted histones. B, HeLa cells growing on a slide chamber were transfected with the pCE-MST1-FL or pCE-MST1-NT plasmid (NT). The caspase-3 inhibitor Z-DEVD-fmk (20 μm) was added to the medium immediately after transfection. At 36 h after transfection, cells were fixed with paraformaldehyde, stained for Myc-Tag (MST1, red), γ-H2AX (green), and Hoechst 33342 (blue), and observed by confocal immunofluorescence microscopy. The bar graphs represent the percentage of apoptotic chromatin condensation in transfected cells from three independent experiments (±S.D.). At least 200 cells were counted each time. The asterisk indicates p < 0.05.