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. 2019 Sep 17;9:13462. doi: 10.1038/s41598-019-50058-7

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© The Author(s) 2019

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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TET2 plays a pivotal role in active DNA demethylation of Sry promoter. (a) RNA-seq based gene expression values (TPM) of Sry, Tet1, Tet2, and Tet3 in gonadal somatic cells at the sex-determining period. n = 2. (b) Expression kinetics of Tet1, Tet2, and Tet3 in gonadal somatic cells. mRNAs were collected from XY gonadal somatic cells at the indicated ts stages and introduced into qRT-PCR analysis. mRNA expression levels in gonadal somatic cells at 16–17 ts stage were defined as 1. Data are presented as mean ± SD. * P < 0.05. n ≥ 3. (c and d) DNA methylation levels in the Sry promoter in E11.5 XY gonadal somatic cells of the indicated genotypes were measured by TAB sequencing (c) and bisulfite sequencing. (d) Blue bar shows the average percentage of methylation at individual CpG sites and red bar shows the average percentage of methylation on total CpG sites. P values for bisulfite sequencing were obtained using the non-parametric two-tailed Mann-Whitney U test. (e) Comparison of DNA methylation kinetics in the Sry promoter in develoing gonads. Gonadal somatic cells were purified from XY Tet2Δ/+ and Tet2Δ/Δ embryos at the indicated ts stages and were then used for bisulfite sequencing analysis.