Immunoblotting of Angpt2 polyclonal antibody (Cat # PAB10091) was used at a 1:500 dilution to detect mouse Angpt2 in supernatants of mouse-angiopoietin-expressing endothelial cells. Lane 1 - mock endothelial cells. Lane 2 - mouse Angptl2 (clone 2-9) expressing cells. Lane 3 - mouse Angpt1 (clone 1-15) expressing cells. Lane 4 - mouse Angpt1 (clone 1-8) expressing cells. Lane 5 - wt endothelial cells. Approximately 20 ug of each lysate was loaded for 10% SDS-PAGE. Immunoprecipitation preceded reaction with primary antibody at room temperature for 1 h. After subsquent washing, a 1 : 5000 dilution of HRP conjugated Gt-a-Rabbit IgG preceded color development.