FIG. 5.

Effect of CaMKKβ inhibition on thrombin-induced AMPK activation. HUVEC were preincubated for 30 min with STO-609 at the indicated concentrations. A. Cells were lysed and subjected to immunoblotting using antibodies against phosphorylated AMPKα (threonine 172) and total AMPKα for counterstaining. A typical experiment and the densitometric analysis of four experiments are shown (means ± SEMs). B. AMPK activity was examined in anti-AMPKα1 immune complexes isolated from 100 μg total protein by phosphorylation of the SAMS peptide. AMPK activity is shown as U/mg lysate protein (mean ± SEM, n = 3), where 1 unit equals 1 nmol ³²PO₄ incorporated into SAMS peptide per min. , P < 0.05 versus untreated thrombin-stimulated controls.