Figure 3. FIGURE 3: Snf1 protein abundance, but neither stability nor phosphorylation, is decreased by Ubc1 disruption.

(A) Early logarithmic phase WT and ubc1Δ strains harboring genomic Snf1-GFP were grown in 2% (H) and 0.05% (L) glucose for 1 hour prior to cell lysis, and an equal amount of total protein was loaded in duplicates for Western analysis of total Snf1 (Snf1^tot-GFP) and phosphorylated Snf1 (Snf1^ph-GFP).

(B) WT and ubc1Δ strains harboring genomic Gal83-TAP or Snf4-TAP were treated as in (A) and Western analysis for TAP abundance is shown.

(C) Assessment of Snf1 protein stability over 3 hours in WT and ubc1Δ strains in the presence of cycloheximide, added at time (0), in 2% glucose. Equal cell numbers were removed at the indicated time points with 40 μg protein loaded.

(D) Biological repeat of Snf1-GFP stability (as in B) is shown with 80 μg protein loaded per lane, and additional timepoints.