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. 2016 May 15;30(10):1211–1224. doi: 10.1101/gad.280685.116

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© 2016 Tsabar 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 3. — The effect of ASF1, RTT109, and RTT101 on Rad53 phosphorylation during repair in the two-DSB system. (A) Rad53 Western blot in a strain that carries only the rapidly repairable SSA (tNS024). Samples were run on a 6% SDS-PAGE gel. (B) Rad53 Western blot after induction of an irreparable DSB (JKM179), one ectopic GC (YJK17), and two repairable HO-cleaved DSBs (2-DSB wild type [YFA01], 2-DSB asf1Δ [YFA02], 2-DSB rtt109Δ [YFA06], and 2-DSB rtt109Δ [YLK03]). Rad53 hyperphosphorylation can be seen as higher-migrating forms. Samples were run on a 6% SDS-PAGE gel. (C) PTC2 overexpression rescues the checkpoint arrest in asf1Δ after induction of two DSBs. Single cells were plated on galactose plates, and G2–M-arrested cells (dumbbells) were counted 24 h after induction. n = 50.