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. 2006 Nov 27;203(12):2703–2714. doi: 10.1084/jem.20051730

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Copyright © 2006, The Rockefeller University Press

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Figure 6. — The rescue of endothelial barrier function and the stimulation of VE-cadherin–mediated adhesiveness caused by JAM-C knockdown is dependent on the activity of Rap1. (A) HDMECs were transfected with control siRNA (open bars) or with siRNA against JAM-C (gray bar). Permeability of HDMECs transfected with control siRNA was analyzed in the absence (−) or in the presence of forskolin (10 μM)/rolipram (20 μM), or the Epac activator O-Me-cAMP (200 μM). Permeability is expressed as the percentage of control, defined as permeability of the control siRNA-transfected HDMECs in the absence of any competitor. (B) The adhesion of HDMECs transfected with control siRNA in the absence (open bars), or in the presence of the PKA activator (Sp-8-Br-cAMPS, 200 μM, filled bars), or in the presence of the Epac activator O-Me-cAMP (200 μM, hatched bars), or of HDMECs transfected with siRNA against JAM-C (gray bars) to immobilized Fc–VE-cadherin or Fc–PECAM-1 is demonstrated. Adhesion is shown as the percentage of adherent cells. (C) HDMECs were transfected with control siRNA (open bars), with control siRNA together with siRNA against Rap1 (shaded bars), with siRNA against JAM-C (gray bars), or with siRNA against JAM-C together with siRNA against Rap1 (hatched bars). Permeability of HDMECs was analyzed in the absence (−) or presence of VEGF (50 ng/ml) or histamine (50 μM). Permeability is expressed as the percentage of control, defined as permeability of the control siRNA-transfected HDMECs in the absence of any stimulus. (D) The adhesion of HDMECs transfected with control siRNA or siRNA against JAM-C to immobilized Fc–VE-cadherin or Fc–PECAM-1 is shown in the absence (open bars), or in the presence of the PKA inhibitor (Rp-8-Br-cAMPS, 200 μM, shaded bars), or in the presence of Rap1 knockdown with siRNA against Rap1 (gray bars). Adhesion is shown as the percentage of adherent cells. Data are mean ± SD of a typical experiment; similar results were obtained in three separate experiments.