Fig. 2. FBPase accumulates in the cytosol in the vid22 mutant. (A) The vid22 mutant was glucose starved and then shifted to glucose for t=0 or t=60 minutes prior to harvest. Total lysates were subjected to proteinase K (PK) digestion in the absence or presence of 2% Triton X-100 (TX). (B) Both vid22 and vid24 strains were glucose starved and then shifted to glucose-rich media for 60 minutes. Cells were homogenized and subjected to differential centrifugation. The percentage of FBPase found in each fraction is indicated. (C) Wild-type cells and the vid22 mutant strains were shifted to glucose for 0, 20 and 60 minutes and harvested. Total lysates were immunoblotted with anti-Vid24p antibodies. (D) Wild-type and vid22 mutant strains were shifted to glucose for 30 minutes and harvested. Differential centrifugation was performed. Proteins from the total (T), high-speed supernatant (S) and high-speed pellet (P) fractions were immunoblotted with anti-Vid24p antibodies.