The endothelial barrier separates the inner blood compartment from the surrounding tissues. At the molecular level, adhesion molecules accumulate at the endothelial cell-cell junction and contribute to maintain vascular integrity. An increase in the endothelial permeability is frequently associated with the deregulation of junctional adhesion. Here, we review how to evaluate the in vitro functions of endothelial cell-cell contacts. We focus this chapter on cell imagery and biochemical analysis of VE-cadherin, the main constituent of adherens junction, and we also provide description of endothelial cell models and methods for studying tight junctions.