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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 replication stress. (A) Representative images of combed DNA fibers from primary MM and UF cells pulse-labeled with thymidine analogues IdU (red) and CldU (green). The MED12 mutation status of UF cells [MED12 WT (M12 WT) or MED12-mutant (M12 MT)] is indicated. (B) Top, pulse labeling scheme. Bottom, percentage of singly labeled fibers retaining only first (Idu) or second (CldU) label indicative of stalled or new forks, respectively. (C) Velocity of fork progression as determined from ldU tract lengths (kb/min). (D) Percentage of bidirectional asymmetric forks, represented as percentage of forks relative to total number of forks in the field. Statistical significance was calculated using One-Way ANOVA followed by Tukey’s Post hoc test, ****p ≤ 0.0001, ***p < 0.001; **p < 0.01. ns, not significant.