Overexpressed Cdc18-CFP foci are associated with Rad22-YFP. (A) The pREP3x∷cdc18 plasmid was introduced into yeast cells expressing the Rad22-YFP fusion. Overexpression of the cdc18 gene was induced by growing cells for 23 h in the absence of thiamin, and Rad22-YFP foci were visualized by epifluorescence microscopy. Cells grown in the presence of thiamin are shown as controls. (B) Genomic DNA content of the above cells grown in the presence or the absence of thiamin was analyzed by FACS. (C) Observation of Cdc18-CFP was achieved by introducing pREP5∷cdc18-CFP plasmid in cells described in A and processed similarly. YFP- (a,d,g) and CFP-tagged (b,e,h) proteins were observed in separate channels as described in Methods, and images were merged with Adobe Photoshop (c,f,i). Representative pictures are shown.
