Figure 4.

In vivo angiogenic effects of nonmutant VEGF-D and VEGF-D^V118M. A: Reactivity with anti–phospho-VEGFR-2 antibodies showing enhanced phospho-VEGFR-2 staining in subject (left panel) compared to control lung (right panel). Nuclei are stained with hematoxylin. B: Filter disks loaded with concentrated conditioned medium from non-transfected human embryonic kidney 293 (HEK293) cells or 2.5 μg of either nonmutant or VEGF-D^V118M derived from HEK293 stable cell lines were placed on chick chorioallontoic membranes (CAMs) of 10-day-old chick embryos and collected after 48 hours. Representative photomicrographs of filter disks and the attached CAMs show increased vessel caliber and tortuosity in the presence of VEGF-D^V118M. C: Quantification of the area of blood vessels (left panel), branching points (middle panel), and vessel diameter (right panel). ^∗∗P < 0.01, ^∗∗∗P < 0.001 (by one-way analysis of variance). Scale bars: 100 μm (A); 1 mm (B).