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. 2021 Jan 11;11:278. doi: 10.1038/s41598-020-79488-4

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Evaluation of ovarian fibrosis during reproductive ageing. (A) Representative processed color threshold images of PicroSirius Red (PSR)-stained ovarian tissue sections used to quantify fibrosis in 2, 6, 12 and 18-month-old mice. (B) Higher magnification images from specific regions boxed in ovaries from 2, 6, 12 and 18-month-old mice. Scale bars are 200 μm. (C) Percentage of PSR-positive staining in ovaries from 2, 6, 12 and 18-month-old mice. n = 4–5 per cohort. Gene expression levels of Col1a1 (D) and Col4a1 (E) in ovaries from 2, 6, 12 and 18-month-old mice. n = 4–6 per cohort. (F) Quantification and representative image of ovarian protein levels of collagen I from 2, 6, 12 and 18-month-old mice. n = 4 per cohort. Data are presented as mean ± SEM; C: Kruskal–Wallis test, Dunn’s multiple comparisons test (p > 0.05). (D,E,F): Ordinary One-way ANOVA, Tukey’s multiple comparisons test: a and b are significantly different among groups (p < 0.05). Each dot represents one animal.