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. 2019 Jan 1;33(1-2):75–89. doi: 10.1101/gad.315978.118

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© 2019 Moquin 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 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 5. — ZPET delays the binding of MRE11 and CtIP to chromatin and slows resection. (A) HA-ZPET expression was induced by Dox in U2OS cells as indicated. Cells were treated with CPT for the indicated durations. Cell lysates were analyzed by Western blot using the indicated antibodies. (B) Cells were plated on coverslips, treated with Dox and CPT as indicated, and analyzed by immunofluorescence using the p-RPA32 (S33) antibody. (C) ZPET knockout cells reconstituted with Dox-inducible HA-ZPET were cultured in the presence or absence of Dox. The cells were then treated with 1 µM CPT for the indicated durations, and whole-cell extracts were analyzed by Western blot using the indicated antibodies. (D) Cells were treated as in C and fractionated into chromatin, nuclear-soluble, and cytoplasmic fractions. The levels of the indicated proteins in these fractions were analyzed by Western blot.