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. 2017 Aug 31;7:10272. doi: 10.1038/s41598-017-09880-0

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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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Gonadotropin up-regulates AR content and K63-AR ubiquitination, but down-regulates K48- AR ubiquitination and RNF6 content in the PCOS rat model. (A) After eCG (10IU, i.p.) injection, antral follicles isolated 48 h thereafter from CTL and DHT-treated rats were lysed and AR and RNF6 content were assessed by Western blot. Results are expressed as mean ± SEM (n = 3). Data were analyzed by 2-way ANOVA and Bonferroni post-hoc test. CTL group: [AR: *P < 0.05 (vs. eCG)]. DHT group: [AR: **P < 0.01 (vs. eCG); RNF6: **P < 0.01 (vs. eCG)]. (B) After eCG injection (10 IU, i.p.), antral follicles isolated from CTL and DHT-treated rats 48 h thereafter were lysed and immunoprecipitated using anti-AR, and the precipitate was immunoblotted with anti-K63 and anti-K48 ubiquitin antibody. Results are expressed as mean ± SEM (n = 3) and analyzed by 2-way ANOVA and Bonferroni post-hoc test. In CTL group: [K48-linked AR poly-ubiquitination **P < 0.01 (vs. eCG + )]. In the DHT group: [K48-linked AR poly-ubiquitination **P < 0.01 (vs. eCG + ); K63-linked AR poly-ubiquitination **P < 0.01 (vs. eCG + )].