Skip to main content
. 2016 Sep 8;7(46):74612–74629. doi: 10.18632/oncotarget.11916

Figure 2. Fyn phosphorylates AMPK on tyrosine 436.

Figure 2

A., B. NIH3T3 cells were co-transfected with 3myc-AMPKα2-Flag and Fyn-CA-V5. 48 h later, cells extracts were prepared and immunoprecipitated with a Flag antibody. Immunoblots of lysates (A) and immunoprecipitation samples (B) were performed with the indicated antibodies. Blots are representative of 3 independent experiments. C., D. Forty-eight hours after co-transfection with 3myc-AMPKα2-Flag, Fyn-CA-V5 and pcDNA3-LKB1, HEK293T cells extracts were prepared and immunoprecipitation was performed using a Flag antibody. Immunoblots of lysates (C) and immunoprecipitation samples (D) were performed with the indicated antibodies. These are representative immunoblots independently performed 3 times. E. Signal quantification of tyrosine-phosphorylated AMPKα levels from (D). Statistical examination between each set was performed. F. Purified Human PRKAA2 (100 ng) and either mock or Flag-Fyn-CA (380 ng) were incubated with the kinase buffer and ATP at 30°C for 30 min. Samples were boiled and separated onto a 8% SDS-PAGE gel, followed by immunoblotting with the indicated antibodies. G., H. HeLa cells were co-transfected by either 3myc-AMPKα2 WT or Y436F (YF) with Fyn-CA-V5. Immunoprecipitation was performed using a Flag antibody. Immunoblots of lysates (G) and immunoprecipitation samples (H) were performed with the indicated antibodies. Images are representative of 4 independent experiments.