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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 4. — JAM-C is essential for HDMEC contractility. (A) Representative immunofluorescence of Rhodamine-conjugated phalloidin showing the distribution of F-actin in buffer- or histamine-treated HDMECs transfected with control siRNA or with siRNA against JAM-C. Nuclei are shown by DAPI staining. (B) The F-actin to G-actin ratio of HDMECs transfected with control siRNA or with siRNA against JAM-C in the absence (open bars) or presence of histamine (filled bars) is shown. The F-actin to G-actin ratio is expressed as the percentage of control, defined as the F-actin to G-actin ratio of the control siRNA-transfected HDMECs in the absence of histamine. Data are mean ± SD (n = 3) of three separate experiments. *, P < 0.05; ^#, P < 0.01. (C) The phosphorylation of MLC was analyzed by Western blot with antibody against phospho-MLC or total MLC in HDMECs transfected with control siRNA or with siRNA against JAM-C. A representative immunoblot for phospho-MLC is shown. The insert shows densitometric analysis of phospho-MLC/total MLC in HDMECs transfected with control siRNA or with siRNA against JAM-C. Data are shown relative to control (phospho-MLC/total MLC in HDMECs transfected with control siRNA was set as 1) and are mean ± SD of three separate experiments. (D) MLC phosphorylation was analyzed as in C in HDMECs transfected with control siRNA or with siRNA against JAM-C in the absence (0 min) or presence of histamine for different time points as indicated.