Vasohibin as an endothelium-derived negative feedback regulator of angiogenesis
J. Clin. Invest. Kazuhide Watanabe, et al. 114:898 doi:10.1172/JCI21152 [Go to this article.]

Figure 2
KIAA1036 inhibits angiogenesis. (A) Preparation of KIAA1036 protein. SDS-PAGE/Coomassie brilliant blue staining is shown on the left and Western blotting on the right. Arrows indicate KIAA1036. (B) Effect of KIAA1036 on network formation by ECs. HUVECs were plated on Matrigel in the absence or presence of KIAA1036 (10 nM). (C) Downregulation of KIAA1036 during network formation on Matrigel. HUVECs were plated on Matrigel, and after the indicated period of incubation, total RNA was obtained and real-time RT-PCR was performed. Values are expressed as mean ± SD of 3 samples. (D) Matrigel implantation analysis was performed as described in Methods. The vessel number per low-power field in 3 different fields was counted for each sample. Values are expressed as mean ± SD of 5 animals. (E) Mouse corneal micropocket assay was performed as described in Methods. Arrows indicate the site where pellets were implanted. Neovascular area (mm²) was determined using NIH Image. Values are expressed as mean ± SD of 5 eyes. (F) CAM assay using adenovirus vectors was performed as described in Methods. The nylon mesh containing adenovirus/Matrigel mixture was placed on the peripheral zone of the CAM, where vascular structure would not appear (left panels). Four days after adenovirus infection, vascular formation was evaluated by macroscopic observation (right). Arrowheads indicate the site where the nylon mesh was placed. *P < 0.05, **P < 0.01.