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. 2022 Feb 18;12:826655. doi: 10.3389/fonc.2022.826655

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Copyright © 2022 VanderVere-Carozza, Gavande, Jalal, Pollok, Ekinci, Heyza, Patrick, Masters, Turchi and Pawelczak

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Replication protein A inhibitor (RPAi) impact on replication fork dynamics. (A) Schematic depiction of experimental design. DNA was pulse-labeled with IdU for 20 min. After IdU removal, replication forks were stalled by the addition of HU or left to replicate with vehicle treatment. HU was removed, and replication was labeled with CldU. Following CldU, cells were treated with the DDRi or vehicle. (B) Quantification of results from DNA fiber analysis in H460 cells treated with the indicated agents. HU was used at a final concentration of 2.5 mM, the ATRi VE-822 at 2 µM, and the RPAi 329 at 50 µM. Data presented are combined from three individual experiments (100 fibers analyzed per experiment; 300 fibers total). Red bar indicates the median value of CldU/IdU. Data were analyzed by ANOVA with Bonferroni test for multiple comparisons (****p < 0.0001).