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. 2013 Jul 30;41(19):8943–8958. doi: 10.1093/nar/gkt645

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Mus81-Mms4 is necessary to respond to DNA damage occurring during S-phase. (A) Sensitivity to MMS. Serial dilutions (10-fold) of normalized log-phase cultures were spotted onto YPD plates containing the indicated amounts of MMS and incubated at 30°C for 48–72 h. MUS81⁺MMS4⁺ (W303-1a strain); mus81Δ (YMV39 strain), mms4Δ (YMV48 strain), mus81Δ mms4Δ (YMV49 strain). (B) Cells were synchronized in G1 and released in medium ±MMS (0.033%). The cells were collected at the indicated time points and the DNA content was determined by flow cytometry. (C) Cell viability after MMS treatment during S-phase. The different MMS concentrations used are indicated at the top of each panel. (D) Checkpoint activation. Upper panel: immunoblot analysis of Rad53. Middle panel: in situ autophosphorylation assay for Rad53. Bottom panel: fluorescence microscopy; G1-blocked cells were released into medium with MMS (0.033%) for 2 h and spindle elongation was analysed. Wild type (YJT126 strain); mus81Δ GFP-TUB1 (YMV22 strain); rad53Δ GFP-TUB1 (YJT127 strain).