Redistribution of Rad Proteins Figure 3. Dynamics of bleomycin response during tlc1 mutant outgrowth. (A) Spot test analysis of serial 5-fold dilutions of the indicated strains on YEPD or YEPD containing indicated concentration of bleomycin. tlc1 spore colonies were passaged either by restreaking on solid plates (top) or repeat dilutions in liquid medium (bottom). (B) Southern blot analysis of the XhoI- (top) or four-base cutter (bottom)-digested genomic DNA isolated from the strains in (A) probed with telomere sequences. The numbers in both panels indicate numbers of restreaking or dilutions. The arrows in (A) indicate the rise of type I (top) or type II (bottom) survivors. doi:10.1371/journal.pone.0008224.g003 Figure 4. Recruitments of Rad51 and Rad52 at an HO cut site are reduced in type I survivors. (A) The structure of the URA3 allele on S. cerevisiae chromosome V for these experiments. The position of various oligonucleotide primers (arrows) used for PCR and real-time PCR are shown. (B) ChIPs were performed and quantitated by real-time PCR as described in Materials and Methods. Error bars represent standard deviations for at least three independent experiments and are provided for these strains. Bindings of Rad51 and Rad52 to HOcs in type I cells differed from those in wild-type in a statistically significant manner (P values = 0.0002 and 0.0115, respectively). These differences were not observed when a mock immunoprecipitation was performed with normal rabbit IgG (data not shown). (C) Rad51 and Rad52 expression levels in these strains were analyzed by Western blot analysis. doi:10.1371/journal.pone.0008224.g004 PLoS ONE | www.plosone.org 6 December 2009 | Volume 4 | Issue 12 | e8224