Fig. 4: MeCP2 binding at methylated miRNA loci slows Pol II allowing miRNAs biogenesis.

a Pol II-pSer2 occupancies for each group of miRNAs across 100 bp of miRNA region and 100 bp of the flanking regions. Regions upstream of the 5‘ ends of the pre-miRNAs between all methylated groups vs. the unmethylated groups were tested. All tests were statistically significant. ***p < 0.001; t-test. b Relative Pol II-pSer2 to Pol II-pSer5 (Ser2/Ser5) occupancies analyzed by ChIP over select miRNA regions in WT and TKO mouse ESCs. Immunoprecipitated DNA was quantified by qRT-PCR using primers spanning the indicated pre-miRNA sequences. Data were normalized to input DNA. c MeCP2 occupancies in regions upstream of start sites between all methylated groups vs. the two unmethylated groups were tested. All tests were statistically significant except for the test between the bilateral and the depleted groups. ***p < 0.001; t-test. d Relative MeCP2 occupancy over miRNA genomic regions in WT and TKO ESCs determined by ChIP analysis. Immunoprecipitated DNA was quantified by qRT-PCR using primers spanning the indicated pre-miRNA sequences. Data were normalized to input DNA. e SP1 occupancy for each group of miRNAs. The regions upstream of the start sites between all methylated groups vs. the unmethylated groups were tested. All tests were statistically significant. ***p < 0.001; t-test. f MeCP2 protein levels in WT mouse ESCs upon treatment with siMeCP2 or siControl. GAPDH was used as a loading control. g Efficiency values for the methylated miRNAs and unmethylated miRNAs from WT mouse ESCs upon treatment with siMeCP2 or siControl. Data were normalized to RPLP0 for pri-miRNAs and U6 for mature miRNAs. h Relative Pol II-pSer2 to Pol II-pSer5 (Ser2/Ser5) occupancy analyzed by ChIP in WT mouse ESCs upon treatment with siMeCP2 or siControl. Immunoprecipitated DNA was quantified by qRT-PCR. Data were normalized to input DNA. All error bars represent ± SEM (n = 3); * represents p < 0.05; ** represents p < 0.01; *** represents p < 0.001; NS = not significant; t-test. Source data are provided as a Source Data file.