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. 2019 Oct 1;9:12618. doi: 10.1038/s41598-019-48772-3

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© The Author(s) 2019

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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Difference of tissue-sparing effect between 200, 50, and 12.5 μm-slit radiation. (a) Immunohistochemical images of ex vivo testes culture tissues 1 h after 10 Gy 200, 50 and 12.5 μm-slit X-ray microbeam irradiation. Staining for γ-H2AX, Anti-GENA (anti-germ cell-specific antigen antibody), and Hoechst (for DNA) is shown as red, green, and blue, respectively. Scale bars, 100 μm (upper); 20 μm (middle and lower). (b) Representative images showing Acr-GFP expression in single cultures following 0 Gy (control), 5 Gy 200, 50 and 12.5 μm-slit (50%), and 2.5 Gy uniform X-ray irradiation (100%) obtained at 14, 16, 18, 20 and 22 dpp. Scale bars, 500 μm. (c) Chronological Acr-GFP expression changes after the 5 Gy 200, 50 and 12.5 μm-slit (50%) X-ray irradiation. A minimum of three tissue samples each from different donor mice were used for each experiment. Data represent the mean Acr-GFP expression ± SD. Asterisk indicates P < 0.01.