Epidemiological and histological findings implicate matrix Gla protein in diastolic left ventricular dysfunction
H&E staining (A), Sirius Red staining (B), cMGP (red [D]), CD31 (green [F]), α-smooth muscle actin (αSMA, green [G]). Panels are multiple stains highlighting: (i) cMGP (red), cell membranes (WGA, green) and nuclei (TO-PRO3, blue) in panel C; (ii) cMGP (red) and endothelium (CD31, green) in panel H; (iii) cMGP (red), αSMA (green) and nuclei (TO-PRO3, blue) in panel I; and (iv) a negative control without cMGP and CD31 primary antibodies in panel E. Two sets of magnifications visualize the microvascular endothelium. Images J, K and L correspond to the square in panel H, while images M, N and O are taken from another area of panel H. cMGP is abundant in the small muscularized vessels (panels D, E, F, G, H and I), in microvascular endothelium (panels J, K, L, M, N and O) and in (panel C). The scale bar represents 100 μm in panels A, B and C; 50 μm in panels D, E, F, G, H and I; and 10 μm in panels J, K, L, M, N and O. L indicates vascular lumen.