Extended Data Figure 6. Mechanical force on PlxnD1 results in integrin activation, but ligand stimulation causes ECs to collapse.

(a) BAECs were incubated with anti-PlxnD1-coated beads and subjected to force (10pN) for 5 minutes. ECs were fixed and stained with HUTS4 antibody to mark ligated β1 integrin. Mean fluorescence intensity was quantified using ImageJ software. Values were normalised to the “no force” condition. Location of the beads are highlighted in yellow circles. n=50 cells/condition from 3 independent experiments. The data represent mean±SEM. P-values were obtained by performing two-tailed Student's t test using Graphpad Prism ****p<0.0001, scale bar represents 10 μm. (b) BAECs were incubated with Sema3E or vehicle, fixed and stained with anti-vinculin antibody to mark focal adhesions. Focal adhesion number was quantified using ImageJ software. Values were normalised to the “vehicle” condition. n=30 cells/condition from 3 independent experiments. The data represent mean±SEM. P-values were obtained by performing two-tailed Student's t test using Graphpad Prism ****p<0.0001, scale bar represents 10 μm.