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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 3: — Dissociation of MYPT1 from myosin II during apoptosis. (A) HeLa cells expressing V5-MYPT1-WT were treated with DMSO or TNF/CHX for 3 h. Immunoprecipitates prepared with anti-mouse immunoglobulin G (IgG, lanes 3 and 4) and anti-V5 antibody (V5, lanes 5 and 6), respectively, were immunoblotted with anti–myosin heavy chain IIA antibody (MHCIIA). (B) Pull-down assay from nonapoptotic HeLa cell lysates was performed using FLAG-MYPT1-WT, FLAG-MYPT1-(1-884), FLAG-MYPT1-T696AT853A, FLAG-MYPT1-T696AT853A-(1-884), FLAG-MYPT1-T696DT853D, and FLAG-MYPT1-T696DT853D-(1-884), respectively (lanes 2–7). The precipitates were probed with anti–myosin heavy chain IIA antibody (MHCIIA). Filled and open arrowheads indicate the positions of full-length and caspase-3–cleaved MYPT1, respectively.