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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 (??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 meanąSEM. ns: P???0.05; *P?0.05; ***P?0.001. ??SMA indicates ??smooth muscle actin; DAPI, 4?,6?diamidino?2?phenylindole; NPNT, nephronectin; and VEGF, vascular endothelial growth factor.