In vivo matrigel plug and tumor angiogenesis in 129/Sv mice. (A) From left to right, control, bFGF, bFGF+α3(IV)NC1, VEGF, VEGF+α3(IV)NC1. E indicates ECs; M, matrigel; SM, smooth muscle. Scale bar: 40 μm. Arrows point to the blood vessels. The number of blood vessels in the matrigel plugs was counted in 10 fields at ×200 magnification. Images were viewed using a Zeiss AX10 camera (Carl Zeiss) with a 100×/1.4 NA objective lens, captured using a Flex Digital IEEE-1394 camera (Sheerin Scientific), and processed using SPOT advanced imaging software 3.1.0 (Diagnostic Instruments) and Adobe Photoshop 7.0 (Adobe). (B,C) Number of blood vessels and Hb content quantification from panel A. The mean (± SEM) are shown. *P < .01, VEGF with vs without α3(IV)NC1. **P < .01, bFGF with vs without α3(IV)NC1. (D) The graph of the growth of mice tumors with and without α3(IV)NC1 injections. The results are shown as mean (± SEM). P < .005, tumor mice without α3(IV)NC1 injection as control group. (E) The average tumor weights of different groups shown in panel D. (F) Frozen sections (4-μm) from different tumor tissues were stained with anti-CD31 antibody and the number of CD31-positive blood vessels were counted. Blood vessel quantification results are shown as the mean (± SEM). *P < .005, mice with vs without α3(IV)NC1 treatment. (G) The circulating VEGFR2-positive cells in the blood of tumor-bearing mice, quantification results are shown as the mean (± SEM). *P < .005, mice with vs without α3(IV)NC1 treatment. (H) Frozen sections from different tumor tissues were stained with anti–COX-2 and CD31 antibodies, followed by FITC rhodamine-conjugated secondary antibodies. CD31 and COX-2 merged positive blood vessels were shown (arrow) in 6 fields at 200× magnification. Images were viewed, captured, and processed as described for panel A. Scale bar: 50 μm.