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. 2015 Sep 28;210(7):1117–1131. doi: 10.1083/jcb.201504007

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© 2015 Grimsey et al.

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Figure 10. — PAR1-mediated endothelial barrier disruption requires p38 signaling, NEDD4-2, TAB1 and TAB2. (A) Endothelial cells pretreated with DMSO or 5 µM SB203580 for 20 min were stimulated with 10 nM α-Th, and permeability was determined. The data in bar graphs (mean ± SD [error bars], n = 3) from 30 min is representative of three independent experiments and were analyzed using a Student’s t test (***, P < 0.001). (B) HUVECs pretreated with DMSO or 5 µM SB203580 for 20 min were stimulated with 10 nM α-Th for 10 min. Cells were immunostained for VE-cadherin and gap formation was determined. The data (mean ± SD [error bars], n = 4) were analyzed using a Student’s t test (**, P < 0.01). Bars, 10 µm. (C) Mice received intraperitoneal injection of PBS or 1 µM SB203580 before injection of PBS, 4 ng/µl VEGF, or 1 µg/µl TFLLRN. The data (mean ± SD [error bars], n = 10 mice) from three independent experiments were analyzed using a Student’s t test (***, P < 0.001). Bars, 5 mm. (D) Endothelial cells transfected with ns, NEDD4-2 (N4-2), or TAB1 and TAB2 (T1/T2) siRNA were stimulated with 10 nM α-Th, and permeability was determined. Data (mean ± SD [error bars], n = 3) in bar graphs are representative of three independent experiments at 30 min, and were analyzed using a Student’s t test (***, P < 0.001).