Fig. 5.

ADAM17 activation or deletion of JAM-A/F11R impairs WSS-dependent endothelial cell alignment. Endothelial cell orientation was quantified in vitro in order to assess WSS response as indicated by cell alignment relative to flow direction. HUVECs were transfected with scrambled (Control) siRNA, exposed to recombinant ADAM17 protein, or transfected with JAM-A/F11R siRNA. HUVECs were then subjected to WSS (8 dyne/cm² flow, for 72 h) and orientation distribution frequency percentage relative to flow direction was measured using the ImageJ directionality plugin or angle measurement tool. Cell frequency in different orientation distribution (a–c), cell average orientation degree (d), and representative images (e, f) obtained from 4 independent experiments are shown at 0, 24, 48, and 72 h of culture. Data on panel d are presented as box-and-whisker plots, in which the minimum, the 25th percentile, the median, the 75th percentile, and the maximum are shown, + indicates the mean of values. For statistical analyses, ordinary two-way ANOVA with Sidak’s multiple comparison test was used. *p < 0.05 compared to control siRNA