Fig. 2
From: The autophagy initiator ULK1 sensitizes AMPK to allosteric drugs
Quantitative global and phosphoproteomic analysis uncovers cellular roles for β1-pSer108. a Representative immunoblots for β1/2-dKO iMEFs-expressing β1 mutants S108A or S108E, stimulated with 2 mM phenformin for 1 h. b Workflow showing the stable isotope dimethyl labeling-based quantitative proteomic and phosphoproteomic approach. c Heatmap showing significantly perturbed cellular phosphoproteins and corresponding phosphosites. Red indicates increased, and green decreased, phosphorylation in β1-S108E compared to β1-S108A-expressing cells. Gray indicates missing phosphopeptide in that replicate. d Immunoblot/densitometry analysis confirming increased PAK2-Ser141 phosphorylation in β1-S108E-expressing cells. n = 3, representative immunoblot is shown. Error bars, mean PAK2-Ser141 phosphorylation (arbitrary units) ± s.e.m. Statistical analysis was performed using unpaired two-tailed Student’s t-test. *P < 0.05 indicates significant increase in PAK2-pSer141 in S108E-expressing cells compared to S108A-expressing cells. e ‘‘Cell cycle, connective tissue development and function, cellular movement’’ is one of the top networks associated with changes in phosphoproteome between S108A and S108E-expressing iMEFs, as identified by Ingenuity Pathway Analysis software
