Figure 3.

Ad.sDll4 increased sprouting but reduced branching. (A) HDBECs were co‐cultured on top of fibroblasts in an in vitro angiogenesis assay. They were treated with conditioned media for either Ad.eGFP, Ad.VEGF, Ad.sDll4, or Ad.VEGF + Ad.sDll4 every other day for 2 weeks (n = 3 experimental replicates, experiment was repeated three times). Cells were then fixed and stained for the endothelial junctional marker, VE‐Cadherin (green) and the nuclear marker, DAPI (blue). (B) Both sDll4 and VEGF + sDll4 resulted in a significantly higher tube density (sDll4: P = 0.0004, sDll4 + VEGF: P = 0.0005), than the control eGFP, yet shorter mean tube lengths (C). Treatment with sDll4 resulted in tubes with the most sprouts (16.33 ± 0.677 P = 0.0091); however, VEGF alone and in combination with sDll4 also increased this (D). Additionally, all three treatments increased the density of branches, with VEGF inducing the most and sDll4 alone resulting in slightly less than the combination treatment. One‐way ANOVA with a Bonferroni post‐test, with a 95% confidence interval. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001