Fig 2.

Effects of rho-associated kinase (ROK) inhibition on phosphorylation of regulatory light chain (RLC) in resting hUM and hVM. In-cell Western (ICW) assays using specific Ab measured monophospho-S19-RLC (pRLC, dashed lines), diphospho-T18/S19-RLC (ppRLC, solid lines) or total-phospho-S19-RLC [p19RLC (pRLC + ppRLC)]. The first data point in all graphs corresponds to the vehicle control (V, 100%). The specific ROK inhibitor glycyl-H-1152 (g-H) caused a concentration-dependent decrease in phosphorylated RLC in human uterine myocytes (hUM) (A) and human vascular myocytes (hVM) (B). For hUM, n = 4. As for all ICW data, results are reported as means ± SEM of% changes compared to vehicle-treated controls. For hVM, there were three independent experiments using different passages of human coronary artery myocytes from passages two to five. For hUM, the effect of g-H on ppRLC was significantly greater (P < 0.01) than on pRLC, determined by two-way anova. In contrast, for hVM, the effect of g-H was greater on pRLC compared to ppRLC.