Fig. 1. NBD-PC but not NBD-PE is trafficked to the vacuole (v) in yeast. The wild-type diploid strain CRY2 was grown to early log phase in SDC at 30°C. Cells were then labeled with 5 µM DMSO-solubilized lipid for 45 minutes at 30°C. After being washed twice with room temperature SDC, DAPI was added to label nuclear (n) and mitochondrial (m) DNA. Cells were incubated at 30°C for 15 additional minutes, harvested, and washed twice with ice-cold SC-azide prior to analysis and imaging by fluorescence microscopy.