Figure 7.

Effect of mARD1A²²⁵ on neovascularization. A) Western blot analysis of HIF-1α and mARD1A²²⁵ protein expression in transplanted B16F10 (left) and MNK74 (right) tumor cells. B) Western blot analysis of VEGFA protein expression in transplanted B16F10 (left) and MNK74 (right) tumor cells. C) BALB/cSlc-nude mice were injected subcutaneously with MKN74, MKN74-mock, or two MKN74-mARD1A²²⁵ clones, A#3 and A#4. After 3 weeks, formalin-fixed primary tumor sections from each mouse were prepared and stained with an anti-VEGFA antibody. The brown color signifies VEGFA in primary tumors. Magnification ×400. Scale bar = 50 μm. A#3 = MKN74-mARD1A²²⁵ #3; A#4 = MKN74-mARD1A²²⁵ #4. One representative image of five per cell line is shown. D) Anti-CD34 staining of xenograft tumors derived from MKN74, MKN74-mock, and MKN74-mARD1A²²⁵ cells, indicative of endothelial cells (left). Numbers of CD34-positive vessels in primary tumors (right) expressed as means and 95% confidence intervals (error bars). Vessels from four or five fields from each of the five tumors per group were scored for quantification. *P = .043, **P = .021, compared with the number of CD34-positive vessels in tumors derived from MKN74-mock cells, as determined with the two-sided Mann–Whitney U test. Magnification ×200. Scale bar = 100 μm.