Figure 3

Extracted cells transport the vital dye FM to the vacuole membrane. (A-D), wild-type cells were grown in YPKG for 3d (t = 0) or transferred to YPD for 30 min (t = 30). Cells were divided and half of the cells were extracted. Non-extracted and extracted cells at the t = 0 and t = 30 min time points were incubated with or without 2% Triton X-100 (TX) for 30 min followed by incubation with trypan blue for 30 min. Cells were examined using a light microscope. (E-L), wild-type cells were grown in YPKG for 3d (t = 0) or transferred to glucose for 30 min (t = 30) and harvested. Cells were incubated in the absence or presence of Triton X-100 (TX) and incubated with FM for 3 hours. The distribution of FM was observed by fluorescence microscopy in non-extracted t = 0 cells (E), t = 30 cells (G), TX treated t = 0 cells (F), and TX treated t = 30 cells (H). The same amounts of t = 0 and t = 30 min cells were subjected to the extraction procedure and treated in the absence (I and K) or presence of Triton X-100 (J and L). Cells were incubated with FM and the distribution of FM was examined by fluorescence microscopy.