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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 mutation diminishes Sry expression in gonadal somatic cells and enhances sex-reversal in XY Jmjd1aΔ/Δ mice. (a) Comparison of Sry mRNA levels between genotypes at the sex-determining period. Gonadal somatic cells were purified from E11.5 embryos of the indicated genotypes and were then used for qRT-PCR analysis. mRNA expression levels in Tet2Δ/+ gonadal somatic cells were defined as 1. Data are presented as mean ± SD. *P < 0.05, **P < 0.01. n ≥ 3. (b) Evaluation of sex development of Tet2-deficient E13.5 gonads by immunofluorescence analysis using antibodies against SOX9 and FOXL2. SOX9 and FOXL2 are markers for testicular Sertoli cells and ovarian somatic cells, respectively. The enlarged box demonstrates testicular tubule-like structures (left). The ratio of SOX9-positive cells to FOXL2-positive cells is summarized in the right. Numbers of embryos examined are shown above the bars. Data are presented as mean ± SD. (c) qRT-PCR analysis of Sry in XY Jmjd1a-deficient gonads and Jmjd1a/Tet2-deficient gonads at E11.5. Each of the samples included one pair of gonads/mesonephros. mRNA expression levels in Jmjd1a-deficient gonads were defined as 1. Data are presented as mean ± SD. *P < 0.05. n = 4. (d) Sex development of E13.5 embryonic gonads of the indicated genotypes was evaluated as in Fig. 4b. The ratio of SOX9-positive cells to FOXL2-positive cells is summarized in the right. Numbers of embryos examined are shown above the bars. Data are presented as mean ± SD. *P < 0.05.