Figure 15. Constitutive downregulation of Wisp-1 functionally reduces endothelial cell network density.

(A) HCAECs were transduced with lentiviral particles containing short hairpin–nontargeting, scramble control plasmids (sh-Ctrl), or Wisp-1 knockdown plasmids (sh-Wisp-1). (B) Clones with the most robust suppression of Wisp-1 were used to assess network density abilities. Thirty thousand cells from each experimental group were seeded in triplicate in 48-well plates on growth factor–reduced Matrigel. In parallel, cells were also treated with TNF-α (10 ng/ml). Density of network branching were determined by software analyses of images (5/well) after 8 hours. Graphs show branching density determined by software analyses. A is an analysis of Western blots from the same experimental group, repeated 3 times. B experiments were performed in triplicate with an n = 15 per group (average density of 5 photographs per group). Results depicted as mean ± SEM of AUs *P ≤ 0.05. P values obtained by 1-way ANOVA with Bonferroni’s post test.