FIGURE 5:

Flow promotes Trio immobilization at junction regions and Trio colocalization with VE-cadherin. (A) Left, ECs were transfected with GFP-TrioFL and subjected to flow (12 h) or left untreated. Flow induces colocalization of GFP-TrioFL (green) with VE-cadherin (white). Red, F-actin. Right, fluorescence intensity of the bar (6 μm in length) on the main figure, showing increased colocalization between TrioFL (green) and VE-cadherin (red). Bar, 25 μm. (B) Pixel overlap between VE-cadherin and GFP-TrioFL under static conditions or flow (12 h) conditions. **p < 0.01. (C) FRAP was performed on GFP-Trio under static or flow (12 h) conditions at junctional regions or cytosolic areas, as indicated. Flow increases the immobile fraction of Trio at junctions, whereas no difference was detected in the cytosol. VE-cadherin–GFP FRAP analysis showed no difference in mobility under static and flow conditions. Data are mean of three independent experiments ± SEM. ***p < 0.001. ns, not significant.