Effect of FXIa on EC permeability on HAECs. (A) HAECs were incubated with vehicle or FXIa for 6 hours in the absence or presence of the VEGFR2 kinase activity inhibitor, SU1498 (5 μM), the ADAM10 inhibitor, GI254023X (10 μM), or RAP (50 ng/mL). Cells were lysed and analyzed by western blotting using an anti–C-terminal VE-cadherin antibody. Results are representative of 3 experiments. Significance (∗P < .05) determined by Kruskal-Wallis testing with Dunns correction for multiple comparisons. (B) HAECs were grown to confluence on gelatin-coated Transwell filters and incubated with FXIa (30 nM) for 6 hours in the absence or presence of PP2 (10 μM), SU1498 (5 μM), or GI254023X (10 μM). Permeability of Evans Blue-BSA was measured after 60 minutes of incubation. Data are mean ± SEM (n = 4). Significance (∗P < .05) determined by Kruskal-Wallis testing with Dunns correction for multiple comparisons. (C) HAECs were incubated with FXIa (30 nM) for 15 to 60 minutes. Cells were lysed and immunoblotted with antibodies for phosphorylated Dab1 Y₂₀₂₀, Src Y₄₁₆, VEGFR2 Y1175, ERK1/2 T₂₀₂/Y₂₀₄, eNOS S₁₁₇₇, PLCγ1 Y₇₈₃, or tubulin. Results are representative of 3 experiments. Significance (∗P < .05) determined by Kruskal-Wallis testing with Dunns correction for multiple comparisons. Data are mean ± SEM (n = 3). (D) HAECs were incubated with FXIa (30 nM) for 15 to 60 minutes in the presence or absence of PP2 or SU1498. Cells were lysed and immunoblotted with antibodies for phosphorylated Dab1 Y₂₀₂₀, PLCγ1 Y₇₈₃, or tubulin. Results are representative of 3 experiments. Significance (∗P < .05) determined by Kruskal-Wallis testing with Dunns correction for multiple comparisons. Data are mean ± SEM (n = 3).