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. 2022 Oct 6;13:5883. doi: 10.1038/s41467-022-32915-8

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© The Author(s) 2022

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. 10 — a Schematic representation of the mouse models used. Bone tissue was collected at 11 weeks; cKO, conditional knockout. b Prrx1 driven TET1 and TET2 knockdown efficiencies in femurs of Tet1/2 double knockout mice. c 5hmC dot blot and relative quantification. d–f hMeDIP-Seq of femurs from Tet1/2 double knockout mice. d Principal component (PC) analysis and 5hmC peaks around selected bone specific genes (e). f Correlation between 5hmC occupancy (TSS + /−30 kb) and gene expression as measured by RNA-Seq; FC, fold change. g Expression of Bone60 genes in conditional Tet1, Tet2 or Tet1/2 femurs versus Ctrl femurs. h Wordcloud representing phenotypes associated with distal intergenic loci with the strongest 5hmC loss in double knockout vs Ctrl femurs as analyzed by GREAT analysis; FDR, false discovery rate. i µCT analysis of L5 spine and relative quantification; TV total volume, BV bone volume, Tb.N. trabecular number, Tb.Th. trabecular thickness, Tb.Sp. trabecular separation (j). Bar graphs represent mean ± SD; *p < 0.05; **p < 0.01; ***p < 0.001. One-way ANOVA analysis with Tukey’s (c) and Dunnett’s (j) multiple comparison tests. FDR-adjusted two-tailed, unpaired t tests in g, two-sided Fishers exact test (f). In c, Ctrl vs Tet1/2 cKO ***p = 0.0009, Tet1 cKO vs Tet1/2 cKO ***p = < 0.0001, Tet1 cKO vs Tet2 cKO **p = 0.005, Tet2 cKO vs Tet1/2 cKO *p = 0.0118. N = 5 (c) & n = 6 per group in j from biologically independent animals. Source data are provided as a Source Data File.