Figure 2.

Fig4 localizes to the limiting membrane of the vacuole. Wild-type cells expressing (A) Fig4-GFP or (B) fig4Δ761–879-GFP were labeled with the fluorescent dye FM4-64 for 10 min at 26°C. The dye was washed away and the cells were chased for 60 min at 26°C. The cells were concentrated and chilled on ice, and the localization of FM4-64 and Fig4-GFP was compared by fluorescent microscopy.