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. 2023 May 22;14:2922. doi: 10.1038/s41467-023-38528-z

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© The Author(s) 2023

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. 5 — a, b Representative images of IMGZ-derived E7.5 (a) and E8.5 (b) embryos carrying Tet1/2/3;Dnmt1−4KO, Tet1/2/3;Dnmt3a/3b−5KO, or Tet1/2/3;Dnmt1/3a/3b−6KO. Green fluorescence indicates the expression of Oct4-EGFP. Three independent embryos were analyzed for each group. Scale bars, 200 μm in (a) and 500 μm in (b). c Frequencies of 5mC and 5hmC modified nucleotides in the genomic DNA of control, Tet1/2/3;Dnmt1−4KO, Tet1/2/3;Dnmt3a/3b−5KO, and Tet1/2/3;Dnmt1/3a/3b−6KO embryos (E8.5) were determined by quantitative mass spectrometry. Data are mean ± s.e.m of three biological replicates. P-values were determined by Student’s unpaired two-sided t-test. d Heatmap shows the similarity of WGBS data between different mutant embryos. The similarity between samples was defined as min-max normalized Euclidean distance reduced by 1. e Bar plots represent the average CG methylation level of whole embryos (E7.5) with different mutations measured by WGBS. Data are mean ± s.e.m of four biological replicates. P-values were determined by Student’s unpaired two-sided t-test. f Summary of phenotypes observed in E7.5 and E8.5 embryos carrying different mutant Dnmt and Tet genes generated by IMGZ. Detailed data of homozygous mutants are presented in Supplementary Data 6. Source data are provided as a Source Data file.