FIG. 3.

Colocalization of Rad51 foci at the RTS1 RFB. (A) Schematic of the recombination substrate plus the lacO array integrated at the ade6 locus on chromosome 3. Asterisks indicate point mutations in the ade6 alleles. (B) Representative spread nuclei from srs2Δ mutant cells containing RTS1 (orientation 2), the adjacent lacO array, and pREP81-LacI-ECFP. The cells were grown to logarithmic phase in EMMG supplemented with thiamine. Spreads were stained with antibodies against Rad51 (red) and the ECFP tag on LacI (green). The DNA is stained with DAPI (blue). Two merged images are shown; the one on the left is an example in which foci are not colocalizing, and the one on the right is one in which they are. (C) Mean number of Rad51 foci per cell in strains FO1791, FO1792, MCW2349, MCW2351, MCW2366, MCW2368, MCW2406, and MCW2408. (D) The percentage of Rad51 foci shown in panel C that colocalize with the lacO array. The data shown in panels C and D are mean values from three independent experiments in which a total of 150 spread nuclei (50 per experiment) were assessed for each strain. The error bars are the standard deviations. The one-tailed Fisher exact test was used to obtain the P values for the indicated comparisons. wt, wild type.