FIG 5.

CreA phosphomutations affect protein accumulation. Strains were grown in minimal medium (MM) supplemented with 1% (wt/vol) xylose for 24 h before 2% (wt/vol) glucose was added for 30 min. Total cellular proteins were extracted, and CreA-GFP was immunoprecipitated. Samples were run on a protein gel and Western blotting was carried out (top). The red arrow indicates full-length CreA-GFP in the presence of xylose (a) and xylose and glucose (b). The bottom shows the total protein input used for immunoprecipitation as stained with Coomassie blue. Furthermore, Western blotting of total protein extract for β-actin antibody was performed and used for normalization. (c and d) Graphs representing densitometry scans of the Western blots described in panels a and b, respectively. Densitometry values for CreA-GFP were normalized by the input. Standard deviations represent the averages from three biological replicates (shown as orange symbols). *, P < 0.05; **, P < 0.01; ***, P < 0.001 in a two-way ANOVA multiple-comparison test using the WT as a reference for each condition.