Fig. 3. Localization of Chs4p590-GFP. (A,B) Chs4p-GFP and Chs4p590-GFP were expressed in the indicated strains, all of them W303 derivatives. Cells of the wild type (A) are shown at higher magnification to show details. Note the absence of plasma membrane staining in all Chs4590-GFP-containing cells. Arrowhead indicates neck localization in the double mutant. Note the high levels of fluorescence in the vacuole. Numbers in each picture indicate the percentage (n>300) of cells showing defined staining at the neck or at the bud. (C) Subcellular fractionations comparing Chs4p and Chs4p590 association to particulate (Po) and soluble (So) fractions with and without 4% urea treatment. The figure represents a western blot analysis using the 3xHA tagged version of each protein. Numbers below bands indicate the percentage of the band in each fraction after quantitative densitometry.