Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Nov 6;11:5643. doi: 10.1038/s41467-020-19516-z

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 3 — a Co-localization events in S-phase cells in indicated conditions and strains, as described on Fig. 2a. p value was calculated by Fisher’s exact test. b MSD of the RFB in OFF and ON conditions in n S phase cells of slx8-29 mutant grown at permissive (25^oC, left panel) and restrictive (32 °C, right panel) temperature over indicated time interval (Δt). p value was calculated as a one sided t-test based on MSD curves. Black bars correspond to SEM. c Diagram of constructs containing the reporter gene ura4-sd20 (green) associated (t-ura4sd20 < ori) or not (t-ura4sd20-ori) to the RFB. The non-functional ura4-sd20 allele, containing a 20-nt duplication flanked by micro-homology, is located downstream of the RFB. Upon activation of the RFB, a restarted fork can replicate the ura4-sd20 and the HR-mediated non-processive DNA synthesis favors the deletion of the duplication, resulting in a functional ura4⁺ gene, generating Ura⁺ cells. As control, the construct devoid of RFB is used to monitor the spontaneous frequency of RS that is then subtracted to obtain the frequency of RFB-induced RS. d Frequency of RFB-induced Ura⁺ reversion in indicated strains and conditions. Each dot represents one sample from n independent biological replicate. Bars indicate mean values ± SD. p value was calculated by two-sided t-test. e Binding of Rad51 to the RFB in WT and slx8-29 strains at indicated temperature. ChIP-qPCR results are presented as RFB ON/OFF ratio for each mutant. Distances from the RFB are presented in bp. Values are mean from three independent biological replicates ± SEM. f Top panel: Scheme of replication intermediates (RI) analyzed by neutral-neutral 2DGE of the AseI restriction fragment in RFB OFF and ON conditions. Partial restriction digestion caused by psoralen-crosslinks results in a secondary arc indicated on scheme by blue dashed lines. Bottom panels: Representative RI analysis in indicated strains and conditions. The ura4 gene was used as probe. Numbers indicate the percentage of forks blocked by the RFB ± SD. g Quantification of resected forks. Values are mean of two independent biological repeats.