Figure 6. The effects of phosphorylation defective mutants of MLC on the thrombin-induced barrier disruption in PAECs.

(a) A representative immunoblot (IB) and a summary (n = 4) showing the relative expression levels of endogenous and exogenous MLC in PAECs, expressing wild type (WT) or phosphorylation defective mutants (T18A + S19A, S19A, T18A) of MLC, or LacZ. Control indicates PAECs without adenoviral infection. (b) Representative immunoblots (n = 4) obtained for the analysis of the phosphorylation of endogenous and exogenous MLC with the indicated primary antibodies, before (−) and 3 min after (+) stimulation with 1 u ml⁻¹ thrombin. (c) A summary (n = 7–10) of the temporal changes in the TEER induced by 1 u ml⁻¹ thrombin in PAECs expressing exogenous MLC or LacZ, as indicated. The data are the means ± SEM. *P < 0.05 vs. WT, according to an ANOVA followed by Dunnett’s post-hoc test.