Fig 3. MDC-labelled vesicles are induced by rapamycin and glucose deprivation in yeast cells.
Pb18 yeasts growing in SD medium for 4–5 days were washed and incubated (1x106 cell ml-1) in Synthetic Dextrose medium as control (SD), or in SD medium containing 0.2 μg ml-1 rapamycin (SD+R); SD medium containing 0.2% glucose (SD0.2%); SD medium without glucose (SD-G). After 2 and 24 hours at 36°C, cells were incubated with MDC at 50 μM for 15 minutes, washed three times in PBS pH 7.2 and then immediately analyzed by fluorescence microscopy as described in Materials and Methods. Autophagic vesicles are stained in green. Arrow indicates yeasts entirely full of buds. Scale bar at 10 μm.