Fig. 3.

Fre8-dTom is in the vacuolar system. The indicated strains were induced to form hyphae in IMDM (A, C, D, left and middle panels) or in serum (B). D, right panel, cells were not induced. Cells were analyzed by fluorescence and differential interference contrast microscopy as described in the Experimental procedures section, and overlay images are shown. C, cells were exposed to the vacuolar dye CMAC in the last 30 min of hyphal formation as described in the Experimental procedures section. Fluorescent microscopy studies of Fre8-dTom localization with IMDM and serum-induced germ tubes are representative of four and two experimental trials, respectively. Experiments with CMAC staining (C) are representative of three cultures over two experimental trials. The following strains were utilized: fre8Δ and fre8Δ-F8T as described in Figure 1; dTom is SC5314-expressing isolated dTomato. The bar represents 5 microns. CMAC, 7-amino-4-chloromethylcoumarin; Fre8-dTom, Fre8 to dTomato; IMDM, Iscove's modified Dulbecco's medium.