FIGURE 1.

N-WASP promotes the formation of AJs. A, B, D, and E, HPAEC transfected with N-WASP (siN-WASP) or scrambled (siSc) siRNA were fixed at the indicated time points after stimulation without (A and B) or with thrombin (D and E). The cells were stained with rhodamine phalloidin or anti-p120-catenin antibody followed by Alexa Fluor-labeled secondary antibodies and visualized by confocal microscope. Scale bars, 10 μm. Inset, immunoblot shows the knockdown of N-WASP at indicated times. Actin was used as loading control. Efficiency of knockdown was quantified using densitometric analysis. B and E, the plot shows mean ± S.D. of interendothelial gaps in siSc and siN-WASP transfected cells at the indicated time points from three individual experiments. * indicates significant increase in the gap area compared to scrambled siRNA transfected cells (p < 0.05); # indicates significant increase in the gap area in monolayers transfected with siN-WASP when compared with monolayers transfected with siSc at each time point after thrombin stimulation (p < 0.05). C and F, HPAEC plated on gold electrodes were transfected with siSc or siN-WASP, and changes in TER were determined in real time at the indicated time points. Data represent mean ± S.D. from three experiments performed in triplicates. * indicates significance from siSc transfected monolayer (p < 0.05). Ω indicates ohms.