Fig. 2

Shear stress potentiates activation and consequent nuclear translocation of NOTCH1 protein. a Endothelial cells (HAECs) were plated to confluency on a y-slide (ibidi) and subjected to laminar flow (9 mL min⁻¹ applied). b MRI flow mapping of y-slide in the presence of flow-conditioned HAECs (48 h) generated flow vector plots across all regions of the y-slide (n = 5). c Flow vector plot of high flow speed (26 dynes cm⁻²) region and corresponding HAEC morphology. Notice cell shape defined by β-catenin expression at cell–cell borders (green) and polarized NOTCH1 (red) protein. Scale bar = 20 µm. d Flow vector plot of low shear stress (10 dynes cm⁻²) region and corresponding HAEC morphology with β-catenin (green) and NOTCH1 (red). Scale bar = 20 µm. e Staining for NOTCH1 (red) in high-flow region compared to low-flow region. Dashed white ovals reveal increase of nuclear NOTCH1 protein under high flow compared to low flow. Scale bar = 20 µm. f Flow vector plot of the back end of the y-slide for nine regions of interest. NOTCH1 (green) nuclear staining was quantified for each of the nine regions and plotted as a function of the measured wall shear stress. For each measurement, ~100 cells were evaluated using n = 5 biological replicates, scale bar = 20 µm