Fig. 8

Pericyte Nck1/2 deletion improves OIR revascularization. a Strategy to assess neoangiogenesis after OIR. Nck2 deletion is induced at P12, P13, and P14. b Flat mounts of P17 Nck1−/−Nck2iMC and littermate control OIR retinas. Insets show avascular area measured for quantification. Lower panels show higher-magnification images of pathological vascular tufts. c Avascular area (top) and NVT (bottom) quantification. Number of retinas used for quantification is indicated. Error bars represent s.e.m. ^⋆⋆⋆P < 0.001, Mann–Whitney U test. d (Left) Quantification of α-SMA (top) and PDGFRβ (bottom) coverage of IB4+ vascular tufts. Number of retinas used for quantification is indicated. Error bars represent s.e.m. NS: non-significant. ^⋆P < 0.05, Mann–Whitney U test. (Right) IB4, PDGFRβ, and α-SMA triple staining of the NVTs. Note the smaller and sometimes α-SMA-negative tufts in Nck1−/−Nck2iMC double-mutant mice (arrows). e (Left) Quantification of DESMIN coverage of IB4+ vascular sprouts in P17 OIR retinas (n = 9 imaging fields for Nck1−/−Nck2l/l from 6 retinas and n = 12 for Nck1−/−Nck2iMC from 6 retinas). (Right) IB4 and DESMIN double staining of the angiogenic sprouts. Error bars represent s.e.m. ^⋆⋆⋆P < 0.001, Mann–Whitney U test. f Quantification of α-SMA coverage of IB4+ vascular sprouts in P17 OIR retinas (n = 10 imaging fields for Nck1−/−Nck2l/l from 2 retinas and n = 23 for Nck1-/-Nck2iMC from 4 retinas). Error bars represent s.e.m. ^⋆⋆⋆P < 0.001, Mann–Whitney U test. g, h Images (g) and quantification (h) of FIBRINOGEN leakage in Nck1−/−Nck2iMC compared to control littermate mice. Number of mice used for quantification is indicated. Error bars represent s.e.m. ^⋆⋆P < 0.01, Mann–Whitney U test. Scale bars, 1 mm (b top), 100 μm (b bottom, g), and 30 μm (d, e)