FIGURE 3.

Elevated levels of Rad52-YFP foci in TORC2-Gad8 mutants. A and B, spontaneous Rad52-YFP foci occur in Δtor1, Δste20, or Δgad8 cells. Cells were cultured in YE liquid medium at 30 °C until mid-log phase, stained with Hoechst and Calcofluor, and visualized live by confocal microscopy. The numbers represent the mean values of three independent experiments, with error bars representing the S.D. Bar, 20 μm. C, quantification of Rad52-YFP foci in nuclei representing different phases of the cell cycle as estimated from cellular and nuclear morphology. D, the assembly and disassembly of Rad52-YFP foci during a transient exposure to CPT. Cells were grown to mid-log phase and arrested in CPT. After 3 h, cells were collected and resuspended in fresh YE medium. Cells were photographed live, at the indicated time points, and the Rad52-YFP foci were counted as described above.