Vascular distribution in the skin flap of VASH2 knockout mice. VASH2 knockout mice were applied to the model of subcutaneous angiogenesis. (A) Immunostaining of CD31 (red) and αSMA (green) in the area 2 to 4 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) Immunostaining of CD11b (red) in the area 0 to 2 mm from the necrotic edge is shown in wild-type and VASH2^−/− mice. Scale bars are 200 μm. (D) Adenoviral-mediated gene transfer was performed to supplement the deficient protein in VASH2 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.