Figure 8.

Rapamycin blocks r-protein gene expression in the absence of ongoing protein synthesis. Cells were grown in SC ethanol to 0.4 OD₆₀₀/ml, the culture was subdivided, and cells were treated with either rapamycin, cycloheximide, or both as indicated for 10 min. Glucose was then added to a final concentration of 2% (wt/vol) as indicated, and cells were incubated for an additional 30 min and then harvested (this time of incubation corresponded to the peak induction of r-protein gene expression observed during glucose upshift [see Figure 7A, lane 3]). RNA was then isolated and analyzed by Northern blotting.