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. 2018 Jun 15;8:9202. doi: 10.1038/s41598-018-27379-0

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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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TUNEL staining of testes sections from 8 weeks old WT and Dcaf17^−/− mice to assess germ cell apoptosis. Testes sections of 8 weeks old WT (A–C) and Dcaf17^−/− (D–F) mice were subjected to TUNEL assay, which detects fragmented DNA in the apoptotic germ cells (green). WT testis section (A–C) shows fewer TUNEL-positive cells (green) (B,C) compare to Dcaf17^−/− testis section (E,F). Image H is positive control for TUNEL assay where the testis section was treated with DNaseI enzyme to generate fragmented genomic DNA. Image K is a negative control where testis section was treated only with labelling solution without terminal transferase. Images A,D,G and J are bright field images of respective TUNEL stained fluorescence images B, E, H, and K. Images C,F,I,L are merged images of respective bright field and fluorescence images. Magnification 200X. Scale Bar (J): 50 µm. The number of TUNEL-positive cells (green) per tubule in the testes sections of WT and Dcaf17^−/− mice aged 8 weeks and 8 months were plotted in the graph (M). The Histogram (M) represents mean ± SEM of 3 animals analyzed for TUNEL assay from each strain. Asterisks indicate statistical significance (p < 0.001 at 8 weeks and P < 0.01 at 8 month). Data were analyzed on Graphpad prism 5 using two-way ANOVA technique followed by post hoc Bonferroni’s multiple comparison test.