FIG. 1.

Effect of thrombin on AMPK activation in endothelial cells. HUVEC were stimulated with different thrombin concentrations for 1 min (A) or with 1 U/ml thrombin for the indicated times (B and C). A and B. Cell lysates were subjected to Western blot analysis using an antibody against AMPKα phosphorylated at threonine 172 or an anti-AMPKα antibody for counterstaining. Representative blots and densitometric analyses are shown (mean ± SEM, n = 3). C. AMPK was immunoprecipitated from 100 μg total protein using an anti-AMPKβ antibody, and the activity of the immune complexes was measured using the SAMS peptide assay. 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 controls.