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. 2020 Mar 4;8:103. doi: 10.3389/fcell.2020.00103

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Copyright © 2020 Yu, Hui, Kao, Liao, Yang, Hou, Hsieh, Chang, Tsai, Pinskaya, Yang, Chen, Morillon, Tsai and Lin.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Ectopic DNMT3L pulse restored H3K27me3 on the promoter of PRC2-target genes derepressed in old MEFs. (A) An RT-qPCR analysis (±SEM) demonstrated the relative expression of representative genes. The gene expression levels were normalized by Rplp0, and three technical repeats were performed. The y-axis represents the expression fold-changes relative to the gene expression in old MEFs. Representative genes: Aplp1, Dnmajc6, Kcnj4, and Sim2. (B) Anti-H3K27me3 ChIP-qPCR was performed to demonstrate the enrichment of H3K27me3 on the promoter or exon1 of the represented genes. Negative control: mouse IgG. The RNA expression of representative genes was reciprocally correlated to the accumulation of H3K27me3 in the promoter. (C) The occupancy of DNMT3L on the promoter or exon 1 of the represented genes during DNMT3L expression was assessed by ChIP-qPCR with an anti-FLAG antibody. RFP-expressing MEFs served as DNMT3L-negative controls. Mouse IgG was used as the negative control for ChIP.