Effect of COX-2 inhibition on Wnt/β-catenin activation by vGPCR. (A) HUVECs expressing vGPCR were transiently transfected with TOPflash and pRL-TK plasmids. Twenty-four hr after transfection, the cells were treated with 100 μM of NS-398 or vehicle control for 24 hr. Cell lysates were collected and analyzed for luciferase expression. Data are presented as the mean from two independent experiments each performed in duplicate. ns – not significant. (B) HUVECS expressing vGPCR were pretreated with 100 μM NS-398 or vehicle control for 24 hr and seeded onto Matrigel-coated wells in medium supplemented with 100 μM NS398 or vehicle control for 6 hr. (C) The number of branch points was determined and expressed as the average from three replicate wells. ns – not significant. (D) Cyclin D1, VEGFA and MMP-9 mRNA expression was measured by qRT-PCR in control (BABE), vGPCR- expressing cells, and vGPCR-expressing cells that were treated with 100 μM NS-398 for 24 hr. The data are presented as fold change (mean +/− SEM) relative to control (BABE) sample. *p < 0.05; **p < 0.01; ns – not significant.