The spatiotemporal expression profile of VASH2. (A) Immunostaining of VASH2, CD11b, and F4/80 in the area 0 to 2 mm from the necrotic edge. Scale bars are 200 μm. (B) Total RNA was isolated from each area of the skin flap. Quantitative real-time RT-PCR was performed to show mRNA levels of VASH2 in each area. Each value was standardized with β-actin. (C) The basal level of VASH2 mRNA in HUVECs or THP-1 cells was determined by RT-PCR. (D) After confirming bone marrow reconstitution, the subcutaneous angiogenesis experiment was performed. Immunostaining of VASH2 in the area 0 to 2 mm from the necrotic edge is shown. Arrowheads indicate GFP-positive and VASH2-positive cells. Scale bar is 50 μm.