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Fig. 1 | Epigenetics & Chromatin

Fig. 1

From: Widespread recovery of methylation at gametic imprints in hypomethylated mouse stem cells following rescue with DNMT3A2

Fig. 1

Five canonical imprinted regions in mouse and validation of gDMR methylation assays in WT ESCs. a Schematic showing the main features of the imprinted domains examined, along with positioning of imprinted gDMRs (narrow rectangles): (i) H19/Igf2 region; (ii) Igf2r region; (iii) KvDMR/Kcnq1 region; (iv) Snrpn; and (v) Peg1. Exons are represented by dark-filled boxes, and expression of the genes from maternal or paternal alleles is indicated by bent arrows above or below the line, respectively. Wavy lines and narrow bars below the line represent long non-coding RNA and CpG islands (CGIs), respectively. The number of CpG sites (circles) covered by pyrosequencing or clonal analysis, and enzyme restriction site positions and fragment sizes for COBRA are all shown below the gDMRs. Filled circles represent methylated sites, while empty circles are unmethylated. CCCTC-binding factor (CTCF) binds to H19/Igf2 ICR on the maternal chromosome only (i) to block access of the downstream enhancers to Igf2. On the paternal chromosome, CTCF binding is blocked by methylation and the enhancers preferentially interact with Igf2 rather than H19. b Methylation level of imprinted gDMRs in various mouse tissues and WT ESCs was quantified by pyrosequencing. Sperm was used as a control: all maternally imprinted genes exhibited low levels (<25%) of methylation and the paternally imprinted gene H19 showed high levels (>75%) of methylation here, as expected. All maternally and paternally imprinted genes show ~50% methylation at gDMRs in lung, heart and brain as assessed by pyrosequencing. In WT ESC, methylation levels were very comparable, with only Igf2r a little high. Error bars indicate s.e.m. c Clonal analysis of Snrpn and H19 gDMRs in parental WT ESCs. Each circle represents a CpG site assessed by bisulfite clonal analysis. Percentage total methylation is indicated at the bottom of each panel

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