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. 2018 Sep 1;64(6):511–522. doi: 10.1262/jrd.2018-093

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©2018 Society for Reproduction and Development

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)

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Fig. 4. — Analysis of acyline-treated mouse testes. (A, B) Real-time PCR (A; n = 6) and immunostaining analyses (B) of acyline- and leuprolide-treated testis. Recipient mice were treated with busulfan to remove endogenous spermatogenesis 1 month before analysis. (C) Quantification of tubules with CLDN5⁺ Sertoli cells. Cells in 10 tubules were counted. (D) Functional assessment of the BTB in acyline-treated mouse testis. Recipient mice were injected interstitially with biotin (green). (E) Real-time PCR analysis of busulfan-treated testes that received lentivirus that overexpress Cldn5 (n = 3). Samples were recovered 2 days after microinjection. (F) Macroscopic appearance of recipient testis that received GS cell transplantation. (G) Colony counts (n = 18 for Cldn5; n = 16 for control). Bar = 20 μm (B), 50 μm (D), 1 mm (F) . Counterstain: Hoechst 33342 (B, D). Asterisk indicates statistical significance (P < 0.05).