Fig. 9

Nephronectin promotes intercapillary network formation and smooth muscle cell recruitment. A, Phase‐contrast images of aortic rings at 7 days in culture. Aortic rings were embedded in collagen‐I solution and cultured in supplemented Opti‐MEM culture medium with or without VEGF or/and recombinant mouse NPNT in 96‐well cell culture plates. B–D, Quantitative analysis of maximum distance covered by angiogenic sprouts (n=7) (B), periaortic vascular capillary branch points density (n=4) (C), and the maximum diameter of the periaortic vessels (n=9) (D). E and F, Representative examples of the appearance of pericytes/smooth muscle cells (α‐smooth muscle Actin‐positive, arrows) surrounding vascular sprouts (E) and quantitative analysis (F). Statistical significance in (B) was determined by 1‐way ANOVA followed by a pos thoc test and in (C–F) by a 2‐tailed Student's t test. Data are mean±SEM. ns: P ≥ 0.05; *P≤0.05; ***P≤0.001. α‐SMA indicates α‐smooth muscle actin; DAPI, 4′,6‐diamidino‐2‐phenylindole; NPNT, nephronectin; and VEGF, vascular endothelial growth factor.