Fig. 7

NCK promotes pericyte migration and retinal vascularization. a Experimental strategy to delete Nck1 and Nck2 in pericytes during retinal development (P0–P5). The red triangles indicate tamoxifen injections. b Western blot against NCK2, PDGFRβ, and GAPDH using P5 total lung lysate from P0/1/2 tamoxifen-injected mice. c IB4, NG2 (left), and IB4, DESMIN (middle) double staining of the angiogenic front and IB4, DESMIN double staining of the plexus (right) of P5 retina. d Quantification of NG2 (left) and DESMIN (right) coverage of IB4+ vessels of the angiogenic front and the plexus of P5 retina. Note the specific decrease of pericyte coverage in the angiogenic front of Nck1−/−Nck2iMC double-mutant mice. Number of retinas used for quantification is indicated. Error bars represent s.e.m. ^⋆⋆⋆P < 0.001, Mann–Whitney U test. e (Left) IB4, NG2, and α-SMA triple staining of P5 retina. (Right) Higher magnification of the boxed areas. f Quantification of α-SMA coverage. Number of retinas used for quantification is indicated. Error bars represent s.e.m. g IB4 staining of retinal flat mounts of P5 mice with the indicated genotypes (negative images of the fluorescent signal). h Quantification of branchpoints, vascular area, and vascular outgrowth. Number of retinas used for quantification is indicated. Error bars represent s.e.m. ^⋆⋆P < 0.01, ^⋆⋆⋆⋆P < 0.001, Mann–Whitney U test. i IB4, ERG1/2/3 double staining of the angiogenic front of P5 retina with the indicated genotype. j Quantification the ERG1/2/3-positive EC. Number of retinas used for quantification is indicated. Error bars represent s.e.m. NS: non-significant. Mann–Whitney U test. k Quantification of sprouts in the angiogenic front. Number of retinas used for quantification is indicated. Error bars represent s.e.m. ^⋆⋆P < 0.01, Mann–Whitney U test. Scale bars, 30 μm (c, e, i) and 500 μm (g)