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. 2018 Dec 3;8:17531. doi: 10.1038/s41598-018-35719-3

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© The Author(s) 2018

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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Analysis of macrophages in testis and epididymis: Immunoperoxidase technique using F4/80 antibody to detect macrophages was applied to cryostat testis (A,B) and epididymis (C,D) sections from WT mice, or DKO mice from Group 1 (DKO1) or Group 2 (DKO2). Similar results were observed between WT and DKO1 mice. Representative images of WT mice (A,C) show scarce number of F4/80+ cells scattered in the interstitium. In contrast, DKO2 mice (B,D) show numerous F4/80+ cells in the testis interstitium (B, arrow) close to a damaged seminiferous tubule (B, asterisk) and in the lumen of epididymal tubules with spermatozoa heads within the macrophage cytoplasm (D, arrow). DKO2 correspond to the enlarged organs from either unilateral- or bilaterally affected mice.