Fig. 2: NMVs preferentially adhere to atheroprone regions in vivo.

Fluorescently labelled NMVs (green) were injected via the tail vein into ApoE^−/− mice that had been fed a Western diet for 6 weeks. After 2 h, mice were culled and en face immunostaining of the mouse aortic arch was performed. Representative en face images of NMV adhesion in atheroprotected (outer curvature, a, b) and atheroprone (inner curvature c, d) regions of the aorta, visualised by confocal fluorescence microscopy. Endothelial cells were identified by staining with anti-CD31 antibody (red) and cell nuclei were identified using TO-PRO Iodide (magenta). Outer and inner curvature of the ascending aorta were identified by anatomical landmarks and confirmed by characterising the phenotype of endothelial cells; those at the outer curvature were aligned, elongated and uniform—a characteristic of cells under high shear, whereas cells in the inner curvature had a disorganised appearance. Samples were visualised using a ×100 objective at ×1 zoom (a, c) and at ×6 zoom (b, d). Scale bars = 20 µm (a, c), 5 µm (b, d). e Quantification of adherent NMVs presented as mean ± SEM (n = 4 animals) and statistical significance evaluated using a paired t-test test. *P < 0.05. Source data are provided as a Source Data file.