Extended Data Figure 8. Endothelial cell alignment to shear stress lacks dependency on nitric oxide but is coupled to calpain.

a, Frequency of HUVEC alignment induced on the orbital shaker. Data for each test condition were normalized to their own control. Test conditions were 0.3 mM L-NMMA (n=3) and transfection with eNOS siRNA (eNOS.si.1) compared with mock transfection (n=4). b, Protein abundances from mass spectrometry analysis for the indicated proteins in 3 Piezo1−/− relative to 3 Piezo1+/+ E10.5 embryos. Calpain-2 and its substrates were less in Piezo1−/− embryos. The effects were relatively specific because more than 1300 of the detected proteins were unchanged by Piezo1 depletion; data for 2 examples (myosin light chain-3 and integrin-β1) are shown. c, Fluorescence images of Piezo1-GFP in a HUVEC before (upper image) and after (lower image) 15 dyn.cm⁻² for 50 min. The small solid arrows point to focal adhesion structures at the trailing edge of the cell. Scale bar, 10 μm. Representative of n=4. Error bars are s.e.m.