Fig. 10. Vps20p and Vta1p are required for transport of FM4-64 from late endosomes to the vacuole. Wild-type (SF838-9D), vps20 (SF838-9Dvpl10) and vta1 (PLY3046) cells were grown to early exponential phase and then incubated with 2 µM FM4-64 at 0°C for 30 minutes. Cells were washed in ice-cold YPUAD and resuspended in YPUAD at 30°C without FM4-64 (0'). Cell aliquots were removed at the indicated times after shift to 30°C (5', 10', etc.), washed in 1% sodium azide, 1% sodium fluoride, 100 mM Tris pH 8.0, and viewed by fluorescence and DIC microscopy. Bar, 5 µm.