Figure 4.

De novo synthesis is not required for Nce102 internalization. Subcellular distribution of Nce102-GFP was checked in cells (strain Y240) prior to the onset of Nce102 internalization (left column), and following the subsequent cultivation without (middle) or with 100 µg/mL cycloheximide (Sigma Aldrich, St. Louis, MO, USA). Independent of the presence of the drug, significant increase in the GFP signal at the vacuolar membranes can be recognized after 3 h of cultivation. GFP fluorescence signal on transversal optical sections (upper row) and brightfield images (lower) are presented. Inverted fluorescence intensities in false-coloured lookup table are shown to maximize contrast. Vacuoles are denoted by asterisks, cell nuclei by arrowheads. Bar: 5 μm.