Fig. 6. Both monomethyl and dimethyl M-C6-NBD-PE are trafficked to the
vacuole in yeast. The wild-type diploid strain CRY3 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 and mitochondrial
DNA. Cells were incubated at 30°C for 15 additional minutes, harvested and
washed twice with ice-cold SCNaN3 prior to analysis and imaging by
fluorescence microscopy.
