Fig. 4. Fur4p-GFP is retained in the class E compartment in vps27^ts mutant cells. Parental and vps27^ts mutant cells transformed with pFL38gF-GFP were grown to exponential growth phase at 24°C in galactose medium. Cells were transferred to 37°C for two hours. The synthesis of Fur4p-GFP was stopped by adding glucose and incubating for 10 minutes at the same restrictive temperature. Cycloheximide (100 µg/ml) was then added. (A) Uracil uptake (permease activity) was measured at 37°C at various times after the addition of cycloheximide. The results are expressed as a percentage of the initial activity. (B) Nomarski optics and distribution of Fur4p-GFP fluorescence in parental and vps27^ts mutant cells. Cells were observed two hours after adding cycloheximide at 37°C. Cells were viewed by Normarski optics or with a FITC filter set.