Figure 2.

Rescue of VEGF-mediated permeability by activating Tie2 pathway comparable to dexamethasone in porcine brain endothelial cells in vitro. (a) Primary porcine brain endothelial cells isolated and cultured as described previously⁷¹ were subjected to TEER measurements in cellZscope device (Nanoanalytics).⁸⁰ Expression of EC marker (VE-Cadherin/green, Claudin-5/red, DAPI/blue) indicate purity. VEGF (10 ng/ml), DEX (100 nM) and AKB 9785 (1 µm) treatments were performed in basal serum-free MCDB-131 medium upon reaching a plateau in TEER indicating confluency and barrier maturation. (b) Dexamethasone treatment resulted in high TEER values almost three-fold compared to controls and furthermore was able to rescue VEGF-mediated BBB breakdown. (c) AKB 9785 treatment was also able to tighten the BBB to similar levels as DEX and rescued VEGF-mediated permeability in this model.