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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 3. — JAM-C mediates an increase in paracellular permeability. (A) The expression of JAM-C, ZO-1, and actin was analyzed by Western blot in HDMECs transfected with siRNA against JAM-C or with control siRNA as indicated. (B) The expression of JAM-C in HDMECs transfected with siRNA against JAM-C was 32 ± 7% of the expression of JAM-C in HDMECs transfected with control siRNA. Data are mean ± SD of six separate experiments. (C) HDMECs were incubated for 1 h without (100% control) or with VEGF (50 ng/ml) or histamine (50 μM) in the absence or presence of Fc (open bars) or Fc–JAM-C (gray bars) (each at 20 μg/ml). Permeability for FITC-conjugated dextran is expressed as the percentage of control, defined as permeability in the absence of any stimulus or competitor. (D) HDMECs transfected with control siRNA (open bars) or with siRNA against JAM-C (gray bars) were incubated for 1 h without (−) or with 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. Data in C and D are mean ± SD (n = 3) of one experiment typical of three separate experiments. *, P < 0.05; ns, not significant. (E) Representative immunofluorescence of histamine-treated HDMECs transfected with control siRNA or histamine-treated HDMECs transfected with siRNA against JAM-C, as indicated, showing the distribution of JAM-C and ZO-1. Note the more contiguous lining of interendothelial contacts in HDMECs upon JAM-C knockdown.