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. 2022 Apr 13;12:6169. doi: 10.1038/s41598-022-10188-x

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© The Author(s) 2022

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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MED12-mutant UFs are characterized by aberrant R-loop accrual and activated replication stress signaling. (A and B) Representative IHC staining with (A) R-loop specific S9.6 and (B) pATR (Ser428) specific antibodies. Shown are images from 3 patient-matched tissue sample sets (PT1-PT3), each set comprising MM, MED12-wild-type (M12 WT) and MED12-mutant (M12 MT) UF tissue. (C) Quantified expression of S9.6 and phosphorylated (activated) replication stress signaling markers pATR (Ser428), pRPA (Ser33), pCHK1 (Ser317), pCDC25A (Ser124), and pATM (s1981). Data were quantified from IHC analysis of 10 patient-matched tissue sample sets. Stained sections were scanned using an Aperio ScanScope® CS system, and individual nuclei as well as nuclear and cytoplasmic marker localization were identified and quantified using Aperio ImageScope software. Signals were normalized to the number of nuclei in each section. Statistical significance was calculated using One-Way ANOVA followed by Tukey’s Post hoc test, **p ≤ 0.01; *p ≤ 0.05.