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. 2018 Aug 14;37(18):e95329. doi: 10.15252/embj.201695329

Figure 4. RMER4B sequence in the Rasgrf1 DMR is essential for de novo DNA methylation of the Rasgrf1 DMR .

Figure 4

  1. DNA methylation status of a portion of the Rasgrf1 differentially methylated region (DMR) in 0–2 dpp Mili +/− and Mili −/− prospermatogonia. Regions corresponding to B, C, D, E, and F are demethylated in Mili −/− prospermatogonia. Vertical bars represent DNA methylation levels at CpG sites determined by whole‐genome bisulfite sequencing (see below). The top dark gray bar represents a portion of the Rasgrf1 gametic DMR (Tomizawa et al, 2011). The CpG cluster is shown. The deleted RMER4B sequence is indicated.
  2. Deletion of key sequences (circles labeled as ①–③) in the Rasgrf1 DMR and chr7 piRNA cluster. Shown is our proposed mechanism in which piRNAs generated from RMER4B sequence in chr7 piRNA cluster target RMER4B sequence in pit‐RNA for the establishment of DNA methylation of the Rasgrf1 DMR.
  3. DNA methylation status of the Rasgrf1 DMR in Rasgrf1 ΔRMER4B/ΔRMER4B and control spermatogonia is shown. The regions examined are shown in (A). Black (white) circles represent methylated (unmethylated) CpGs.
  4. Rasgrf1 RMER4B functions in cis. Wild‐type and mutant alleles of the Rasgrf1 DMR in Rasgrf1 +/ΔRMER4B spermatogonia were analyzed separately.