Figure 6: TET2 is mainly responsible for the 5mC–5hmC conversion in the HOXA cluster during RA induction.
From: Hydroxylation of 5-methylcytosine by TET2 maintains the active state of the mammalian HOXA cluster
The histograms show the distribution of 5mC and 5hmC at four regions of the HOXA cluster in NT2 cells transfected with control siRNAs (non-targeting pool—kd scrm), siRNA pools against TET1 (kd T1), TET2 (kd T2), TET3 (kd T3) and TET1/TET2 (kd T1/T2). NT2 cells were treated for 3 days with RA, replated and transfected with siRNAs. After 3 additional days, cells were harvested and genomic DNA for (h)MeDIP analysis with antibodies specific for 5mC and 5hmC was isolated. Immunoprecipitated DNA was amplified by gene-specific q-PCR. The following regions were analysed: (a) HOXA1 promoter/first exon, (b) HOXA2 promoter/first exon, (c) HOXA3 second CGI and (d) HOXA4 second exon. Enrichments were calculated relative to the unmethylated UEB2B control. 5mC values are shown as black bars and 5hmC values as grey bars. Histograms show the results of three independent experiments. s.d.'s are indicated by error bars.
