H2A.B facilitates transcription elongation at the methylated DMR. (A) Down-regulation of H2A.B does not affect the Pol II deposition at the TSS of Kcnq1 and Igf2r genes. ChIP analyses in the TSS regions (region B of the Kcnq1 gene and region A of the Igf2r gene) are performed using an anti-RNA Pol II antibody. Data are presented as mean ± SEM (n = 3). (B) Down-regulation of H2A.B arrests the pSer2 Pol II at the DMR of Kcnq1and Igf2r. ChIP analyses at the DMR regions (region C of the Kcnq1 gene and region B of the Igf2r gene) are performed using an anti-RNA Pol II pSer2-CTD antibody. Data are presented as mean ± SEM (n = 3). (C) Based on the SNPs between mus and cas, allele-specific deposition of pSer2 Pol II in Kcnq1 and Igf2r is determined by restriction enzyme BmgBI digestion or SSCP. (D) Down-regulation of H2A.B by shRNA does not affect the DNA methylation status at the DMR of Kcnq1; down-regulation of H2A.B by shRNA does not affect the DNA methylation status at the DMR of Igf2r after RA treatment.
