Skip to main content
Figure 6 | Genome Biology

Figure 6

From: Redistribution of H3K27me3 upon DNA hypomethylation results in de-repression of Polycomb target genes

Figure 6

Demethylation with a DNA methyltransferase inhibitor results in redistribution of H3K27me3. (A) Global quantification of the 5mC base by HPLC. Values are 5 mC as a percentage of total C in Dnmt1+/+ MEFs treated with vector control or 5-aza-dC. Error bars represent standard error of the mean (SEM). **P <0.001 by t-test. (B) ChIP-qPCR for H3K27me3 in Dnmt1+/+ MEFs treated with vector control or 5-aza-dC using primers for the promoters of the indicated genes. Enrichment for each immunoprecipitation (IP) is expressed as percentage of DNA in input. Non-specific IgG IP and the Actb promoter are shown as negative controls and the Chdh promoter is shown as a positive control. Error bars indicate ±SEM of two experiments. (C, D) Boxplots showing the relative expression (log2) of the indicated gene set, and all genes, between vector control and treatment with 5-aza-dC in Dnmt1+/+ MEFs. The gene sets compared are those that are both H3K27me3 down and >1.5 fold upregulated at mRNA level in Dnmt1-/- MEFs (C), and H3K27me3 up in Dnmt1-/- MEFs (D). **P <0.005 by Wilcoxon rank sum test relative to all genes. (E) qRT-PCR quantifying expression of the indicated genes in Dnmt1+/+ MEFs treated with vector control or 5-aza-dC. Expression is normalized to Actb and expressed relative to vector control. Error bars represent ±SEM of three replicates. 5-aza-dC, 5-aza-2-deoxycytidine; DMSO, dimethyl sulfoxide; IgG, immunoglobulin G.

Back to article page