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. 2021 Jan 15;10:581459. doi: 10.3389/fonc.2020.581459

Figure 4.

Figure 4

HEYL expression in endothelial cells promotes endothelial cell invasion. (A) Retinal blood vessel growth in neonatal (day 6) retina in HeyL+/+ mice extends farther than in HeyL-/- mice. The blue lines in the figure indicate the extent of outgrowth of the blood vessels while the red arrows define the length achieved in retina of wild type mice. (B) A significant difference is observed in the length of the blood vessels from center to periphery of the retina of 6-day old HeyL+/+ and Heyl-/- mouse pups. (C) Matrigel invasion assay of HUVEC-HEYL or HUVEC-GFP cells. (D) Quantification of the number of the invaded cells in a Matrigel invasion assay shows HUVEC-HEYL cells migrate 2X faster than control HUVEC–GFP cells. (E) (Left panel) 3-D fibrin gel invasion assay was used to measure growth of sprouts of HUVEC-GFP or HUVEC-HEYL cells at day 1, 3 and 5. Right panel: The average sprout length was measured in the two groups. Quantification of three determinations is shown. (F) Matrigel plugs containing bFGF examined 10 days after sc implantation in wild type HeyL+/+ and HeyL -/- mice show different extent of mouse blood vessel infiltration (n=5 mice per group). (G) Sections of matrigel from (F) were stained with Masson’s Trichrome to visualize ECM surrounding blood vessels show greater infiltration in HeyL-/- mice compared to HeyL+/+ mice (n=5 per group). (H) Impaired growth of mouse mammary tumor, E0771, in the mammary fat pad of syngeneic HeyL-/- mice compared to HeyL+/+ mice (n =12 per group). *P < 0.05; **< 0.05; ***< 0.001.