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. 2018 Jan 23;8:1431. doi: 10.1038/s41598-018-19624-3

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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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Expression of purinoceptors P2RX4 and P2RX7 in peritubular cells. (a) Expression of P2RX4 and P2RX7 mRNA was revealed in HTPCs stemming from four individual patients (1–4) and in the human testis (+). Patient-derived HTPCs were additionally screened for the presence of smooth muscle cell markers ACTA2 and CNN1 and absence of the mast cell marker TPSAB1. Negative controls: non-reverse transcription control (−RT), non-template control (−). (b) Immunoblotting confirmed P2RX4 and P2RX7 expression in HTPCs from three different patients (a–c). Note that gel/blot images were cropped and full-length gels/blots are presented in Supplementary Fig. 5. (c) Basal P2RX4 (n = 8) and P2RX7 (n = 8/6) receptor mRNA levels at 6 h and 24 h varied between cells derived from individual patients, but also ACTA2 levels (n = 8) did not remain constant. Data are geometric mean with 95% confidence interval. (d) P2RX4 expression was detected in peritubular cells, germ cells and interstitial tissue, while P2RX7 expression was found in peritubular cells and vessels (not shown) solely. P2RX4 expression in the tubular wall overlapped with SMA and CNN1 expression and absence of tryptase staining in consecutive sections. Insets: Negative controls (pre-adsorption for P2RX4, omission of primary antibody for P2RX7); Bars = 20 µm.