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. 2021 Aug 13;12:4926. doi: 10.1038/s41467-021-25227-w

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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. 2 — a The CENTLEIN protein was completely absent in the Centlein^−/− testis. Immunoblotting of CENTLEIN was performed in the Centlein^+/+, Centlein^+/⁻, and Centlein^−/− testes. GAPDH served as a loading control. Biologically independent mice were examined in three separate experiments with similar results. b Centlein-deficient male mice were completely infertile. The fertility assessment experiments were performed in Centlein^+/+, Centlein^+/⁻, and Centlein^−/− male mice (n = 6 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, ****P < 0.0001. c The size of the testes was not altered in the Centlein^+/+, Centlein^+/⁻, and Centlein^−/− mice. d Quantification ratio of body weight in Centlein^+/+, Centlein^+/⁻, and Centlein^−/− male mice (n = 5 independent experiments). Data are presented as mean ± SEM. e Quantification ratio of testis weight in Centlein^+/+, Centlein^+/⁻, and Centlein^−/− male mice (n = 5 independent experiments). Data are presented as mean ± SEM. f The histomorphology of Centlein-deficient seminiferous tubules was similar to the control groups as shown by H&E staining. L: Leydig cells, Ser (brown): Sertoli cells, P (blue): pachytene spermatocytes, rSt (green): round spermatid, spz (red): spermatozoa. g Quantification ratio of seminiferous tubules with vacuoles in the Centlein^+/+, Centlein^+/−, and Centlein^−/− testes (n = 3 independent experiments). Data are presented as mean ± SEM. h Acrosome and nucleus morphology in different steps of spermatid development was normal in Centlein-deficient mice. The periodic acid-Schiff (PAS) and hematoxylin staining was performed in Centlein^+/+ and Centlein^−/− mouse. i Quantification ratio of spermatids with abnormal acrosome in the Centlein^+/+ and Centlein^−/− testes (n = 3 independent experiments). Data are presented as mean ± SEM. Source data are provided as a Source data file. Blue dots indicate Centlein^+/+mice, green dots indicate Centlein^+/⁻ mice, and red dots indicate Centlein^−/− mice.