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. 2013 Mar 15;24(6):748–756. doi: 10.1091/mbc.E11-08-0740

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© 2013 Iwasaki et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — Effect of MYPT1 depletion on apoptotic cells. (A) HeLa cells were transfected with scrambled siRNA (lane 1), human MYPT1 siRNA (lane 2), or human MYPT1 siRNA and V5-MYPT1-WT vector (lane 3). Forty-eight hours after transfection, the cells were treated with DMSO, TNF/CHX, or TNF/CHX/zVAD-FMK for the indicated times. Total cell lysates were immunoblotted with anti-MYPT1 antibody. The full-length and caspase-3–cleaved MYPT1s are indicated using filled and open arrowheads, respectively. (B) The dead cells in the indicated time were counted after staining the cells with FITC-annexin V and PI. The apoptotic index was calculated by determining the percentage of dead cells in the field (n = 100–200). Results shown are mean ± SD of four independent experiments. Statistical significance was determined using Student's t test (*, p < 0.005; **, p < 0.05).