FIGURE 8:

Glucose starvation–induced expression and surface concentration of Gpa2-GFP. (A) Wild-type cells expressing GFP-tagged Gpa2 were grown in glucose and 90 min raffinose conditions before lysates were made for immunoblot analysis probing with α-GFP and α-GAPDH antibodies. (B) Gpa2-GFP expression levels in glucose and 90 min raffinose treatment were quantified from immunoblots (A) using imageJ and normalized to GAPDH signal. * indicates unpaired t test (p = 0.0287). (C) Wild-type cells expressing endogenously expressed Gpa2-GFP were imaged by Airyscan in glucose (left) and following 2 h raffinose treatment (right) presented as max-intensity projections from four z-stack slices from center-focused cells Arrows showing vacuole (V; yellow) and cytoplasm (C; white) are shown. (D) Mean-intensity measure­ments for vacuole, cytoplasm, and surface from cells in (C) were performed. (E) Max-intensity projections of 3D confocal images of four z-stack slices covering the top-focused area of the cell. (F) Cells exhibiting surface puncta of Gpa2-GFP were quantified as a percentage of total population. * indicates unpaired -test (p < 0.0001). Scale bar, 5 µm.