Impaired VEGF-induced FITC microsphere extravasation in tsad^−/− trachea in the presence of flow and intact pericyte coating. (A) WT and tsad^−/− mice were injected (i.v.) with 30 nm fluorophore-coupled microspheres (green) combined with saline (not depicted) or VEGF. 30 min later, whole-mount preparations of trachea were immunostained with anti-CD31 (red). Bottom panels show magnification of boxed area in top panels. Representative images are shown of three separate experiments using three to four mice/genotype each time. Bars, 100 µm. (B) Quantification of data in A. n = 4 mice/genotype from 1 of 3 experiments. ***, P < 0.001. (C) Quantification of histamine-induced extravasation of microspheres in WT and tsad^−/− trachea. n = 4 mice/genotype from 1 of 3 experiments. (D) WT and tsad^−/− mice were injected (i.v.) with DyLight 594 lectin (red), microspheres (green), and VEGF. 30 min later, whole-mount preparations of trachea were prepared and assessed by confocal imaging. Representative images are shown of two separate experiments using three mice/genotype each time. Bar, 20 µm. (E) Immunofluorescent staining with antibodies for NG2/desmin and CD31. Representative results of two separate experiments using three mice/genotype each time. Bar, 20 µm. (F) Quantification of data from one experiment as shown in E. n = 3 mice/genotype.