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. 2014 Feb 27;10(2):e1004146. doi: 10.1371/journal.pgen.1004146

Figure 5. Cultured AVSMC from mice with combined CathA/Scpep1 deficiency show increased reactivity to ET-1.

Figure 5

A. Pharmacological antagonists of ET receptors, BQ610 and BQ788, reduce MLC-2 phosphorylation in AVSMC treated with ET-1. Cultured AVSMC from WT mice were treated or not for 30 min with 2 µM BQ610 or BQ788 followed by 5 min induction with 100 nM ET-1 as indicated on the figure. Total protein extracts were analyzed by Western blotting using antibodies specific for phosphorylated (pThr18/Ser19-MLC) and total MLC-2 protein. Panel shows representative data of 3 independent experiments. Right panels show ratios (means and S.D.) of signal intensities for phosphorylated and total MLC-2 estimated with ImageQuant software. * p<0.05 in paired two-tailed t-test. B. Increased MLC-2 phosphorylation in AVSMC from double-deficient mice. Cultured AVSMC from WT, Scpep1−/−, CathAS190A and DD mice were treated for 5 min with 100 nM ET-1. Total protein extracts were analyzed by Western blotting using antibodies specific for phosphorylated and total MLC-2 protein. Panel shows representative data of 3 independent experiments. Graph below the panel shows ratios (mean values and S.D.) of signal intensities for phosphorylated and total MLC-2 protein. * p<0.05 in paired two-tailed t-test.