FAJs are tensile junctions. (a) Time-lapse images of HUVECs expressing p120-catenin–mCherry (red) and Vinculin-GFP (green) during treatment with a low dose of Cytochalasin D. Note the specific and rapid displacement of the Vinculin-containing junction, whereas the Vinculin-negative junction does not move in this time period. See corresponding Video 7 for 16-min time-lapse recordings. (b) Quantification of the mean distance of translocation ± SEM of the p120-catenin signal within 60 s after Cytochalasin D treatment of Vinculin-positive junctions (n = 14) and Vinculin-negative junctions (n = 14) from five different experiments as determined by manual measurements in ImageJ. P-value was calculated with a two-tailed, homoscedastic Student’s t test. (c) Time-lapse images of FAJs in HUVECs expressing VE-cadherin–GFP (green) and Lifeact-mCherry (red), before and 15 s after laser ablation at the indicated region. See corresponding Video 8 for an ∼1-min time-lapse recording, which is representative of multiple experiments. The image on the right is a kymograph showing the intensity of VE-cadherin–GFP in time along the dotted line (shown is the maximum intensity pixel of a 10-pixel-wide line). (d and e) IF images of control and thrombin-treated HUVECs stained with the conformation-sensitive α18 rat monoclonal antibody for α-catenin (green), phalloidin for F-actin (blue), and antibodies for VE-cadherin (d) or Vinculin (e; red). Bars: (a) 5 µm; (c) 5 µm; (d and e) 20 µm.