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. 2017 Jun 14;7:3469. doi: 10.1038/s41598-017-02018-2

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© The Author(s) 2017

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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SM increases P450-aromatase expression in Sertoli cells. Sertoli cells were cultured for 48 h at G or in RCCS. (A) Real time PCR of P450-aromatase showing a strong increase of mRNA level under microgravity condition. (B) Real time-PCR for P450-aromatase in Sertoli cells treated or not with 1 mM 8-Br-cAMP. cAMP inducibility is lost under SM. (C) Semiquantitative RT-PCR of P450-aromatase and KL in Sertoli cells treated or not for 48 h with 1 mM 8-Br-cAMP. P450-aromatase expression is stimulated by 8-Br-cAMP at G but not under SM, while KL expression is stimulated by 8-Br-cAMP in both culture conditions. (D) Western blot analysis of P450-aromatase in Sertoli cells after 48 h of culture at G or under SM. Histogram on the right reports the mean of densitometric analysis of four different experiments. Values were normalized by reference to values for actin. (E) Chemiluminescence immunoassay for estrogen level in the culture medium of Sertoli cells. Sertoli cells were cultured in the presence or absence of 50 nM testosterone at G or in SM for 48 h. SM causes an increase of estrogen in the medium when cells are treated with testosterone. At least three different experiments were performed. Bars represent s.d. Asterisks indicate: *P < 0.05 and **P < 0.01.