Fig. 6. CDH5 partnering is shear stress dependent.

COS-7 cell data obtained by FRET/FLIM after expressing PIEZO1-mTurquoise2 with (+) or without (−): a–e PECAM1-SYFP2 (PECAM1); f–j CDH5-mVenus (CDH5). Prior to imaging, cells were preconditioned for 24 h with laminar shear stress (10 dyn.cm⁻²) followed by static condition for 30 min and then 10 min 10 dyn.cm⁻² (shear) or continued static condition (static). For 3 independent experiments each, the box plots show the lifetimes for PIEZO1-mTurquoise2 at cell-cell junctions: e +PECAM1 ***P = 2.64 ×10⁻⁴ shear and ***P = 1.37 ×10⁻⁴ static, both compared with –PECAM1; j +CDH5 ***P = 2.63 ×10⁻⁴ shear and **P = 0.0079 static, both compared with –CDH5. +CDH5 shear cf + CDH5 static **P = 0.00036 and the lifetimes for PIEZO1-mTurquoise2 at non junctional regions: d +PECAM1 **P = 0.0021 shear and *P = 0.011 static, both compared with –PECAM1; i +CDH5 showed no significant differences to the –CDH5 under shear and static conditions. The superimposed data points are for individual junctions (N = 9) and non-junctions (N = 9) from separate images. P values are from Mann–Whitney tests with Bonferroni correction.