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. 2020 Apr 21;10:6751. doi: 10.1038/s41598-020-63290-3

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© The Author(s) 2020

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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Isolation and analysis of Sox3 null adult undifferentiated spermatogonia. (A,B) Flow-sorting strategy for isolation of undifferentiated spermatogonia (EpCAM + c-KIT– α6-integrin+) for gene expression analysis. Percentages of cells within gates are indicated. (C) Graph shows percentage of EpCAM+ germ cells within c-KIT– α6-integrin+ undifferentiated cell gate from flow cytometry analysis of wildtype and Sox3 null adult testis of A and B. Mean values ± s.e.m. are shown (n = 5 wildtype and n = 6 Sox3 null mice). Significance was determined by two-tailed unpaired t-test, P < 0.01 (**). (D) RT-qPCR analysis of sorted undifferentiated spermatogonia (EpCAM + c-KIT– α6-integrin+) from wildtype and Sox3 null adult testis. Expression levels are corrected to β-actin and normalized to a control sample. Mean values ± s.e.m. shown (n = 5 wildtype and n = 6 Sox3 null mice). Significance was determined by two-tailed unpaired t-test and selected P values are indicated, P < 0.01 (**), P < 0.0001 (****).