Figure 1. Norrin-containing conditioned medium promotes endothelial cell growth/motility in an EAhy926 endothelial cell wound healing assay; Norrin blocking antibody inhibits wound healing in monolayer of human endothelial cells EAhy926.

Panel (A) and Panel (B). Wound healing assay demonstrating growth and migration of EAhy926 endothelial cells at time 0 (panel A) and after 24 hours (Panel B). After 24 hours, Norrin blocking antibody (12.5 ug/ml) treated EAhy926 endothelial cells migrate shorter distances than control isotype IgG treated cells. Panel (C). The quantitative results of the migrated distance measurement demonstrate graphically that anti-Norrin blocking antibody impedes endothelial cell growth and migration toward the denuded zone. Panel (D). Norrin mRNA is not expressed basally in the CRC cell line CaCO2. Transfection of these cells with a Norrin/pCMV Script expression construct restores the Norrin mRNA expression. Expression levels in several representative stable transfectant clones are depicted. The increased expression above control pCMV-transfected clones is statistically significant (p < 0.001). Panel (E). Wound healing assay demonstrating that, after 30 hours, Norrin conditioned medium-treated EAhy926 endothelial cells migrate a longer distance than EAhy926 cells treated with control conditioned medium. After 30 hours, no denuded zone remains in the Norrin treated condition. Panel (F). The quantitative results of the experiment in the Panel E is depicted, indicated that Norrin conditioned medium promotes EAhy926 endothelial cell growth and migration.