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. 2016 Dec 28;26(4):753–767. doi: 10.1093/hmg/ddw426

Figure 5.

Figure 5

Bmi1 (PRC1) but not Suz12 (PRC2) is required for copy-number dependent gene repression. A–J. Muscle cells carrying 1, 5, or 12 DUX4p repeats and an EGFP reporter, knockdown for the PRC1 core subunit Bmi1 (siBmi, A–E), for the PRC2 core subunit Suz12 (siSuz12, F–J) or expressing a non-silencing control (Ctrl, A–J) were tested for EGFP expression. For relative quantification of EGFP protein levels among samples, flow cytometry results (A–B, F–G) of EGFP Median Fluorescence Intensity (MFI) (A,F) and of percentage of EGFP+ events (B,G) are shown. For relative quantification of EGFP at the RNA level, RT-qPCR analyses of EGFP (C, H), Gapdh negative control (D, I) and Hoxc13 positive control (E,J) are presented over β-Actin. Results are expressed over Ctrls (A–J). The mean of the signals obtained from five (A,B, F–J) or four (C–E) independent experiments is shown. The error bars represent SEM of independent biological replicates. Asterisks indicate statistical significance (p value) as evaluated by One way ANOVA Bonferroni’s multiple comparison test. ****= P < 0.0001; ***= P < 0.001; **= P < 0.01; ns = non significant.