Figure 4

The Magoh2 promoter is a direct target for Mll2 and a model example of trxG /PcG opposition (part 3). (A) Bisulfite sequencing results of the Magoh2 promoter before (-) and after (4-OHT) addition of tamoxifen for 4 days, followed by a further 8 days of culture shown below a diagram of the promoter, which illustrates the location of the CpG dinucleotides as well as the relevant restriction sites, Southern probe and PCR primers (small arrows either side of the Hpa1 sites) for the results shown in panels (B) and (C). (B) Southern blot to evaluate CpG methylation at the Nae I site of the Magoh2 promoter as illustrated in panel (A). Because the Nae I site contains two CpGs, resistance to restriction can reflect methylation at either or both sites, as illustrated at the right of the panel. The time course was performed on Mll2F/F; Rosa26-CreERT)/+ cells starting with addition of 4-hydroxy tamoxifen. (C) As for panel (B) except the genomic DNA samples were digested with Hpa I before Q-PCR analysis with the primer pair illustrated in panel (A). These data are plotted (red circles, lower plot), as well as quantitation of the Southern blot shown in panel (B; black circles, upper plot).