]>Distinct Effects of Contraction Agonists on the Phosphorylation State of Cofilin in Pulmonary Artery Smooth Muscle : Figure 4
62741.fig.004
Figure 4: Contraction agonists activate LIMK2 in canine PASMCs. Canine PASMCs were incubated with thrombin (Thr), norepinephrine (NE), endothelin-1 (ET1), angiotensin II (AngII), and 5-hydroxytryptamine (5-HT) for 5 min. LIMK2 was immunoprecipitated from total protein homogenates and kinase activity was assayed by the phosphorylation of recombinant CF in the presence of [ 𝛾 - 3 2 𝑃 ]ATP in vitro. Reaction protein was resolved by SDS-PAGE, gels were stained with Coomassie brilliant blue and then exposed on phosphorimaging screens. CF radioactive bands (top inset) were normalized to protein bands (lower inset) and the agonist-mediated activation of LIMK2 was expressed as percentage of the activity of untreated cell controls (Ctr, bar graph). Mean ± SD, (*) 𝑃 < . 0 5 compared to untreated controls, 𝑛 = 3 .