Fig. 3

MTX reduced enrichment of H3K79me2 in Shh and Sufu genes promoters. A Mouse ESCs were treated with MTX (1 μM) for 12 h. Protein was extracted and subjected to western blotting and detected by Dot1L and H3K79me2 antibodies. B F9 cells were treated with MTX (1 μM) for 12 h for immunofluorescence detection. C, D ChIP-Seq H3K79me2 assemble around TSSs. ChIP-Seq: enrichment of H3K79me2 in mESC cells under MTX (1 μM) 12 h. E ChIP-seq: enrichment of H3K79me2 in Shh, Sufu genes in MTX-treated mESC. F ChIP assays were performed using H3K79me2 antibody in MTX-treated mESCs (1 μM, 12 h). Mouse IgG was used as negative control. Enrichment of H3K79me2 in Shh, Sufu genes were detected by RT-qPCR. Data are shown as mean (n = 3). *p < 0.05, by Student’s t-test