Fig. 2

Invasion and angiogenesis of A549-OKS cells on the CAM. a. Macroscopic images of CAM engrafted with A549-GFP (left panel) and A549-OKS cells (right panel) on embryonic development day (EDD) 15. The part surrounded by the yellow dotted line indicates the grafts. b. Microscopic images of CAM engrafted with A549-GFP and A549-OKS cells on EDD 15. Linear blood vessels are distributed on the surface of the A549-GFP graft. A549-OKS cells make thick vessels twist and induce microvessels. Upper and lower panels are captured in 0.67 and 1.5 × magnification. Yellow arrowheads indicate capillary vessels on the transplanted graft. c. A cross-section view of Fig. 2 b. The right panel shows an enlarged image of A549-OKS cells on the CAM. A549-OKS cells invade the CAM, and the surrounding stroma is thickened. The green, blue, and red dotted lines indicate the surface, bottom, and transplanted graft, respectively. Scale bars: 2.5 mm. d. Immunohistochemistry (IHC) to detect Desmin and ASCT2 of CAM with A549-GFP or A549-OKS cells on EDD 15. Desmin-positive cells (shown in red) and ASCT2-positive cells (shown in green) indicate the chick vessels and the transplanted cells, respectively. The grafts are on the membranes in A549-GFP (left panels) and induce a few vessels. The central panel shows that A549-OKS cells with microvessels invade the membrane. The right panel shows cells located away from the mass. Hoechst was used to stain nuclei. Scale bars: 100 μm