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. Author manuscript; available in PMC: 2009 Nov 15.
Published in final edited form as: Exp Cell Res. 2008 Sep 19;314(19):3466–3477. doi: 10.1016/j.yexcr.2008.09.003

Figure 3. Time-dependent effects of CS on thrombin-induced MLC phosphorylation in stretch-exposed EC and in replated CS-preconditioned EC cultures.

Figure 3

A: Human pulmonary EC were subjected to 18% CS for various periods of time followed by thrombin stimulation (0.1 U/ml, 10 min) with continuing CS stimulation. Analysis of MLC phosphorylation was performed using di-phospho-MLC specific antibodies. B: HPAEC exposed to 18 hrs or 72 hrs of 18% CS were replated onto plastic dishes, and thrombin-induced MLC phosphorylation was measured 16 hrs after replating. Quantitative analysis depicts relative levels of phosphorylated target proteins expressed as ratio of phospho-protein to the total protein in the same lysate. In some experiments, the levels of phosphorylated and total target proteins were detected by immunoblotting of two or more identically loaded membranes with corresponding antibody. Results are representative of three independent experiments. C: Comparison of thrombin-induced MLC phosphorylation in static or CS-preconditioned (5% or 18%, 72 hrs) macro- and micro-vascular pulmonary EC. Results are representative of three independent experiments.