FIG. 5.

The effect of Fbh1 overexpression on Rad51, Rad11-GFP, and Rad22-YFP focus formation. (A) Representative spread nuclei from wild-type cells carrying pREP41 or pREP41-fbh1 grown for 5 h in EMMG containing 10 μM CPT. Spreads were stained with antibodies against Rad51 or the tags on Rad11 and Rad22, as indicated. DAPI-stained DNA (blue). (B) Quantification of data shown in panel A. Each value represents the analysis of at least 100 nuclei from at least three different transformants. Rad51 and Rad11 foci were assessed with the same strain (FO1776) and cultures, whereas Rad22 was assessed with strain FO1752. Numbers on top of each bar are the mean number of foci per nucleus and are compared by t testing, where indicated. Note that neither strain FO1752 (containing rad22-YFP) nor FO1776 (containing rad11-GFP) shows any overt growth defects or hypersensitivity to genotoxins, indicating that the tagged forms of Rad22 and Rad11 are functional (data not shown).