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. 2014 Mar 17;204(6):947–963. doi: 10.1083/jcb.201307119

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© 2014 Banerjee et al.

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Figure 8. — CPC activity spreads from centromeres to chromosome arms after nuclear envelope breakdown. (A) Assay scheme. (B) Cells that have endogenous survivin replaced with the survivin^H80A mutant are unable to phosphorylate histone H3Ser10 in the ZM washout assay. HeLa cells stably expressing vector only, survivin-myc (siRNA resistant [res]), and survivin^H80A-myc (siRNA resistant) were treated with survivin siRNA for 48 h followed by 1 h in ZM, MG132, and monastrol. Bar, 2.7 µm. (C) Mean total phospho–histone H3Ser10 intensity at 0 and 15 min in mock-treated (vector [vec.] only) and survivin siRNA-treated vector only (vector only + siRNA) and survivin-myc (wild type [WT] resistant)– and survivin^H80A-myc (H80A resistant)–expressing cells. Error bars show standard deviations. a.u., arbitrary units. (D) Western blot showing Aurora B, survivin-myc, and endogenous survivin levels. Molecular mass markers are given in kilodaltons.