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. 2012 Feb 15;7(2):e31288. doi: 10.1371/journal.pone.0031288

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Srividya et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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WT, yku70, rtt103, mec1 and rtt103mec1 null strains containing RAD53-Myc was treated with MMS for 2 h and anti-Myc western blots were performed. The slower moving fuzzy band indicates phosphorylation of Rad53. Fuzzy bands are visible in all lanes treated with MMS and contain wild type MEC1. mec1 mutants (lanes 9, 11) do not produce phosphorylated Rad53 upon MMS treatment. Wild type sample was loaded in two lanes of the gel for better comparison of the position of the fuzzy band. Same blot was probed with Sir2 antibody to show that separation of proteins was normal and identical in all lanes.