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. 2019 Sep 23;20(11):e47845. doi: 10.15252/embr.201947845

Figure 2. Embryonic and postnatal development in Vegfr2 Y1212F/Y1212F FVB mice.

Figure 2

  • A
    Body weight. P, postnatal day; w, weeks. Error bars: SD. n = 5–12 mice.
  • B, C
    Hindbrain vessel parameters. (B) Vessel density; isolectin B4 vessel area normalized to wild‐type (WT) mean of the litter. (C) EdU incorporation in ERG‐positive ECs normalized to total ERG‐positive ECs. Each dot represents two fields/mouse; error bars: SD; unpaired t‐test, not significant. n = 5–11.
  • D
    P4‐P7 retina vessel morphology. Isolectin B4 (IsoB4) in black; outer 20% of the vascularized area in pink. Scale bar, 250 μm. P6 insets; sprout morphology. Scale bar, 50 μm.
  • E
    Radial outgrowth. Distance between optic nerve and vascular front. P, postnatal day; error bars: SD; 2‐way ANOVA P < 0.0001; Bonferroni's multiple comparison test, *P < 0.05. P4 n = 3–5 retinas, one retina/mouse, P5 n = 4–7, P6 n = 9–9, P7 n = 4–5.
  • F
    Retina vascularized area. P, postnatal day; error bars: SD; 2‐way ANOVA P = 0.0013. P4 n = 7–9 retinas, one retina/mouse, P5 n = 4–9, P6 n = 9–9, P7 n = 4–5.
  • G
    Retinal vessel density in the retinal front. Quantification of the vessel area in the outer 20% of the vasculature (pink in D) (vessel area normalized to vascularized area). P, postnatal day; error bars: SD; 2‐way ANOVA P = 0.0054; Bonferroni's multiple comparison test, *P < 0.05, **P < 0.01. P4 n = 3–5 retinas, one retina/mouse, P5 n = 4–6, P6 n = 3–5, P7 n = 4–5.
  • H, I
    Proliferation of retinal ECs. (H) EdU incorporation in ERG‐positive ECs normalized to total ERG‐positive ECs, at P6. (I) Isolectin B4 (IsoB4; white), ERG (red), EdU (green) and double‐positive cells (yellow). Lower panel shows ERG/EdU merged staining alone with EdU signal masked by ERG signal. Scale bar, 50 μm; error bars: SD; unpaired t‐test, *P < 0.05. n = 5–8 retinas, one retina/mouse.
  • J
    Retina sprout length. Sprout length from tip to base in P6 retinas; error bars: SD; unpaired t‐test, *P < 0.05; n = 3–5 retinas, one retina/mouse.
  • K, L
    Vessel stability. (K) Isolectin B4 (IsoB4, green) and collagen IV (red) in P4 WT and Vegfr2 Y1212F/Y1212F retinas. Arrows: collagen IV‐positive empty sleeves. A: artery. Scale bar, 50 μm. (L) Collagen IV empty sleeves around main arteries in P4–P7 retinas, three fields/retina/mouse. Error bars: SD; 2‐way ANOVA P = 0.0580; Bonferroni's multiple comparison test, **P < 0.01. P4 n = 7–9, P5 n = 6–12, P6 n = 3–5, P7 n = 4.