Figure 7.

FM4-64 staining of endocytic membranes in arf1 ts mutants. Wild-type (YPH499, WT) and arf1 ts mutant cells (arf1-11, arf1-13, and arf1-18) were grown to a logarithmic phase in YPD medium at 23°C, and then half of the culture was shifted to 37°C and incubated for 1 h. Cells were harvested and resuspended at ∼2 × 10⁸/ml in YPD medium prewarmed to 23 or 37°C and incubated for 10 min. FM4-64 was added to the final concentration of 50 μM. After incubation for another 10 min, the cells were harvested and resuspended in fresh prewarmed medium and the incubation continued. Images of the cells were acquired at ∼15–20 min (0 h) and 1 h after the addition of fresh medium and visualized by fluorescence confocal (left) and Nomarski (right) microscopy. Bar, 5 μm.