FIG 7.
Rasip1 and Radil are required for Rap1-induced endothelial barrier function. (A) Endothelial barrier (Rb) of control HUVEC monolayers (siC) and HUVEC monolayers depleted of Radil (siRadil) or Rasip1 (siRasip1), transduced with control lentivirus or Rap1A(V12) containing lentivirus. Different colors represent independent experiments (n > 4). Averages are indicated by the black line. (For the knockdown efficiencies, see Fig. S1A and B in the supplemental material.) (B) Immunofluorescence of HUVECs transfected with control siRNA (siC), siRNA targeting Radil (siRadil), or siRNA targeting Rasip1 (siRasip1). Cells were grown to confluence and either not stimulated or stimulated with 007-AM 15 min prior to fixation. The cells were stained for β-catenin (red) and F-actin (phalloidin, green). The graphs show the relative intensity profiles of fluorescent signal intensities of phalloidin (green) and β-catenin (red) along the line scans depicted in the immunofluorescent images. Gray lines and numbers correlate to where the line scan crosses an adherens junction, as assessed by β-catenin staining. (For the knockdown efficiencies, see Fig. S1C in the supplemental material.)