Fig 8. Silencing β-catenin abrogates the effects of GSK3-β inhibition on lymphatic endothelial cell network formation in vitro.

Lung lymphatic endothelial cells were treated with vehicle (lipofectamine), or transfected with control scrambled siRNA (control siRNA) or β-catenin siRNA for 48h. Equal amounts of protein from whole cell lysates were analyzed by western blotting with antibodies against β-catenin and β-actin (A). LECs were treated with either lipofectamine, control siRNA or β-catenin siRNA for 48h were placed on Matrigel at a density of 1.2X10⁴ cells/well for 16h and then treated with vehicle (DMSO) or SB216763 (1μM). Cells were then labeled with Calcein-AM (8μg/ml). Random 4X images were then obtained (B) and mesh area analyzed with ImageJ Angiogenesis Analyzer plugin (C). Results are expressed as mean ± SEM for one experiment. *P <0.05 by t-test between same group. Experiment was repeated once.