rTMD1 inhibits angiogenesis in vivo. (A-B) Pichia-expressed rTMD1 inhibits angiogenesis in vivo using a murine Matrigel assay. (A) Schematic representation of Matrigel plugs. The magnification bar is 1 mm. (B) Statistical analysis of murine angiogenic assay. The data represent mean ± SD. Similar results were obtained in 3 independent experiments; n represents the number of plugs for each group. ***P < .001 compared with the PBS. ###P < .001 compared with EGF alone. (C-D) rTMD1 inhibits angiogenesis in vivo in a rat corneal micropocket assay. A micropocket in the cornea was surgically generated and implanted with a pellet containing various combinations of 200 ng of EGF and various amounts of Pichia-expressed rTMD1. (C) Corneal photographs were taken 7 days after implantation. Representative figures are shown. (D) Statistical analysis of corneal neovascularization. The data represent mean ± SD. Similar results were obtained in 3 independent experiments; n represents the number of eyes in each group. **P < .01 and ***P < .001 compared with EGF alone. (E-F) rTMD1 inhibits tumor angiogenesis. (E) Lewis lung carcinoma cells (1 × 10⁶) were administered to C57BL/6 mice by subcutaneous dorsal injection. The mice were given daily IP injections of various concentrations of Pichia-expressed rTMD1 for 2 weeks. Representative figures and statistical analyses of tumor mass are shown. ***P < .001 compared with the PBS control. The data represent mean ± SD; n represents the number of mice in each group. (F) Analysis of intratumoral microvessel density (IMD). IMD was detected by CD31 staining. The density of microvessels was quantified in the most vascular areas of the tumor. ***P < .001 compared with the PBS group. The data represent mean ± SD.