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. 2017 Dec 28;10(1):243–256. doi: 10.1016/j.stemcr.2017.11.020

Figure 5.

Figure 5

TRIM28 Is Required to Maintain DNA Methylation at Paternal but Not Maternal ICRs in Primed hESCs

(A and B) TRIM28 binding at the IG-DMR and the H19-ICR relative to the average TRIM28 binding at 29 maternal ICRs as a control. ICR is underlined.

(C) Percent CpG methylation levels at maternal imprinted ICRs in Ctrl and T28KO(U1-9) primed hESCs (n = 29 maternal ICRs), ns, not significant. Error bars represent SD.

(D) Browser view of ATAC-seq (blue), WGBS (green), and TRIM28 ChIP-seq (yellow) at the IG-DMR ICR, the MEG3 promoter, and the H19 ICR.

(E and F) Relative expression using real-time RT-PCR of MEG3 and H19 in T28KO(U1-9) primed hESCs relative to Ctrl. ∗∗∗∗p < 0.0001. Error bars represent SD.

(G–J) ChIP-qPCR of TRIM28 and H3K9me3 at the MEG3 promoter and H19-ICR, in Ctrl and T28KO(U1-9) hESCs.

(K) Sanger sequencing of SNPs in primed Ctrl and T28KO(U1-9) hESCs and naive hESCs as a control for biallelic expression. Arrows indicate two independent H19 SNPs.