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. 2013 Oct 15;27(20):2259–2273. doi: 10.1101/gad.223180.113

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© 2013 Ragland et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

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Figure 3. — RNF4 suppression prevents replication fork collapse in ATR-deficient cells. (A) Quantitative real-time PCR detection of RNF4 mRNA expression 48 h after lentivirus-mediated transduction. The sequences and regions of RNF4 targeted by these shRNAs are distinct (Supplemental Fig. 5). (B) Representative replication restart following RNF4 suppression, as detected by flow cytometry. (C) Quantification of replication restart results in B. (D) Quantification of replication reinitiation on a per-fork basis in RNF4-suppressed cells, as determined by DNA combing. Note that RNF4 suppression permitted significant recovery of replication in previously unrecoverable Atr^Δ/− cells (B,C); this reinitiation occurs at previously active replication forks but not at novel origins (D).