RASIP1 localizes to points of EC-EC contact. (A-F) Immunofluorescence staining of RASIP1 (red) in control (A-C) and RASIP1 shRNA (D-F) infected HUVECs treated with EGTA and cBiMPs, and permeabilized prior to fixation. Peripheral RASIP1 localized to the vicinity of β-catenin+ structures. (G-L) Whole-mount (G-I) or section (J-L) immunofluorescence of DA ECs from ∼E8.5 mouse embryos stained with RASIP1 (G,J, red in I,L) and VE-cadherin (H,K, green in I,L) antibodies. RASIP1 staining is often found at the cell periphery, close to VE-cadherin+ structures. Arrows indicate intracellular VE-cadherin and RASIP1 staining. Scale bars: (A-F) 20 μm; (G-L) 30 μm; (J-L) 10 μm.