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. 2020 Jul 17;10:11913. doi: 10.1038/s41598-020-68639-2

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© The Author(s) 2020

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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Ascites accumulation in vivo correlates with enhanced collagen anisotropy. (A) C57Bl/6 female mice were injected i.p. with 5 × 10⁶ RFP-tagged ID8-Trp53^−/− or ID8-Trp53^−/− BRCA^−/− syngeneic murine OvCa cells as indicated. MicroCT images were procured at time of injection (week 0) and time of sacrifice (week 2 or week 5–6 post-injection, as indicated). (B) The parietal peritoneum was dissected and prepared for combined fluorescence/SHG microscopy as described in Methods. Shown are representative examples from each cohort to show collagen fiber alignment (grey) and metastatic lesions (RFP-tagged cancer cells, red). (C) Collagen anisotropy was quantified using ImageJ. All results are presented as mean ± s.e.m. and P-values were calculated using a Student’s two-tailed t-test.