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. 2016 Jan 15;15(1):95–105. doi: 10.1080/15384101.2015.1121323

Figure 4.

Figure 4.

MMSET is required for normal cell-cycle progression. (A) HeLa cells were treated with the indicated siRNAs and, 48 hours later, cells were fixed and examined for DNA content by flow cytometry. (B) Cell-cycle profiles from A were analyzed and S phase was plotted for each condition. Error bars indicate SEM. *P < 0.05. (C) HCT116 cells stably expressing MMSET shRNA were reconstituted with WT MMSET or the MMSET deletion mutant (648-1356) and the indicated proteins were examined by western blot. (D) A subset of cells from C was examined for DNA content and BrdU incorporation using flow cytometry. (E) HeLa cells were treated with the indicated siRNAs, synchronized at the G1-S border using aphidicolin, and total cell extracts (TCE) or chromatin and non-chromatin enriched fractionations were prepared 48 hours post-transfection. The indicated proteins were examined by western blot. The numbers beneath the blots provide the densitometric ratio of ORC2, MCM2, or MCM7 signal to H3, setting the value for the control group as 100. Data shown are representative (A, C, D, and E) or an average of 3 independent experiments (B).