Fig. 5

Fig. 5

FOXO1 activation is required for VEGFA-induced vessel regression in mouse retina model. a Schematic diagram showing nucleus FOXO1 distribution in retinal vasculature. In WT, nucleus FOXO1 mainly locate in remodeling endothelial plexus, however they are gathering to the angiogenic front of CDS2-deficient endothelium, which could be further increased by excessive VEGFA stimulation. IB4 and FOXO1 co-immunostaining (b) and quantitative analysis (c) on P7 retinas in control and Cds2^iΔEC mice with or w/o recombinant VEGFA stimulation. Yellow arrows indicate nuclear localization of FOXO1. n = 8 mice per group. d Quantification of IB4 and FOXO1 co-staining of P7 retinal vessels in the remodeling plexus area from WT or Cds2^iΔEC embryos with or without ectopic VEGFA injection. n = 5–8 mice per group. Confocal imaging analysis of P7 retinal vessels stained by IB4 (e) and IB4/COL4 (f) from control or VEGFA-injected Cds2^iΔEC mice treated with AS1842856 (FOXO1 inhibitor) or vehicle. Arrows show regressed vessels. g Quantitative analysis on endothelial area, branch points, angiogenic sprouts and COL4⁺/IB4⁻ empty sleeves of P7 retinal vessels in control and VEGFA-injected Cds2^iΔEC mice with or without AS1842856 treatment. n = 6–10 mice per group. Scale bars, 50 μm (b; left panel), 25 μm (b; magnification figure), 200 μm (e) and 100 μm (f). Error bars, mean ± SEM. *P < 0.05; **P < 0.01 ***P < 0.001; ****P < 0.0001; ns, not significant (P ≥ 0.05). See also Supplementary information, Fig. S7