Figure 8.

Polarized localization of Cdc42 and Rho1 is perturbed in slmΔ mutant cells. (A) The distribution of GFP-Rho1 and GFP-Cdc42 in wild-type and slm1-3 slm2Δ mutant cells is shown. Exponential cultures were grown in selective medium at 25°C followed by growth at 38°C for additional 2 h. Expression of GFP-Cdc42 was induced on SD-Leu-Met medium for 1 h at 25°C before shift to nonpermissive temperature. GFP-Cdc42 and GFP-Rho1 localization was visualized by immunofluorescence in living cells mounted in 1% agarose. (B) Approximately 60 small-budded cells expressing GFP-Cdc42 and GFP-Rho1 were scored for polarized distribution of the GFP signal to the bud (black bars) or depolarized localization to the cortex or the cytoplasm (white bars).