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. 2021 Sep 9;12:5341. doi: 10.1038/s41467-021-24894-z

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Overview of the experimental design to identify genes that are downregulated during ESC-to-EB differentiation and bound by RING1B. Representative colony morphologies for ESCs and EBs are also shown. Experiments were repeated independently at least 2 times with the same results. b Identification of genes at which RING1B binding changes during ESC-to-EB transition. The number of genes in each category is shown in the Venn diagram. Genes bound by RING1B are classified into “Group1” (Log2FC[EB/ES] > 1)., “Group2” (−1 ≤ Log2FC[EB/ES] ≤ 1) or “Group3” (Log2FC[EB/ES] < −1), and are indicated by green, blue and yellow, respectively. c Average binding of RING1B in each group in ESCs and EBs. Metaplot views of RING1B binding at TSS ± 5 kb at each gene group are shown. Genes not bound by RING1B in both ESCs and EBs are further divided into CGI + (associated with CGIs) or CGI- (not associated with CGIs). Data from ESCs or EBs are indicated by green or pink lines, respectively. d Binding of RING1B at selected Group1 genes (Klf4, Tbx3, Pdgfa), and Group3 genes (Sox4, Grhl2), in ESCs and EBs. Snapshots of RING1B distributions in the ESCs and EBs are shown. Gene structures and positions of CpG islands (CGI) are indicated at the bottom. e Changes in the expression of genes differentially bound by RING1B during ESC-to-EB differentiation. Violin plot shows of the expression of genes in each group, in ESCs and EBs.