Vascular distribution in the skin flap of VASH1 knockout mice. VASH1 knockout mice were applied to the model of subcutaneous angiogenesis. (A) Immunostaining of CD31 (red) and αSMA (green) in the area 6 to 8 mm from the necrotic edge is shown. Scale bars are 200 μm. (B) The vascular area was determined from 5 different fields in each area. Data are expressed as the means and SDs; *P < .01, **P < .05. (C) Lectin staining (green) shows the perfusion of new vessels in the area 6 to 8 mm from the necrotic edge. The same section was immunostained for CD31 (red). Scale bars are 200 μm. (D) Adenoviral-mediated gene transfer was performed to supplement the deficient protein in VASH1 knockout mice. AdLacZ was use as the control. Immunostaining of CD31 (red) and αSMA (green) in the indicated area of the skin flap is shown. Scale bars are 200 μm.