Fig. 3
Phosphorylation analysis of Snf1 and Gln3 under laboratory conditions. a Immunoblot analysis of the Snf1 phosphorylation in strains BQS252, C9, EC1118 and M2. Cells were grown exponentially in medium with 2% glucose and shifted to derepressing conditions in 0.05% glucose. Proteins were analyzed using the specific anti p-AMPK α (Thr172) antibody, while total Snf1 protein was detected by an anti poly-His antibody. b Gln3 phosphorylation during nutrient starvation. The epitope Myc-tagged versions of GLN3 were changed from SD (0.1% (NH4)2SO4) to minimal medium without ammonium sulfate of 2 mM MSX, or 200 ng/mL of rapamycin were added for 20 min. Glycolytic protein Pgk1 was used as the loading control. c Similar experiment as in panel b, but by testing glucose removal for 15 and 30 min