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. 2017 Jan 17;7:40372. doi: 10.1038/srep40372

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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To determine if the impact of ERR on molecular players associated with spermatogenesis is reflected at the phenotypic level, we analyzed Protamine incorporation into sperm chromatin in control/knockdown males carrying ProtamineB-EGFP and counted the number of mature sperm. Both control and knockdown testes/seminal vesicles displayed sperm labeled with GFP (Green, Panels A and B) suggesting normal protamine incorporation. Further, control males (Panel A) have mature sperm in distended seminal vesicle (SV; indicated by white arrow). However, knockdown males had sperm (in green) scattered all along the globular testicular mass (Panel B, ERR-TRiP, labeled as T). The number of mature sperm in the SV (Panel C) of ERR knockdown males were significantly reduced (**p < 0.01). All these experiments were carried out twice and sperm were counted in tissues twice with a repeatability of index of 92%.