Fig. 1: Ndrg1 deficiency impairs VEGF-A-induced angiogenesis in mice.
a Immunofluorescent images of NDRG1 (green) and CD34 (red) using a specific antibody in five surgically resected breast cancer specimens. ×400 original magnification; scale bar = 10 μm. b We determined Vegfa mRNA expression in mouse cancer B16/BL6, LLC/3LL, and RENCA cells by qRT-PCR (n = 3 per group). We normalized the data by Gapdh expression levels. c Left: Schematic illustration of experimental mouse dorsal air sac assay. Right: We evaluated tumor angiogenesis by RENCA cells using mouse dorsal air sac assays. We determined angiogenic responses by counting numbers of newly formed blood vessels of >3 mm in length (Ndrg1+/+ group, n = 6; Ndrg1−/− group, n = 7). Arrowheads indicate newly formed vessels with characteristic zigzagging lines; scale bar = 2 mm. d, e We induced corneal neovascularization by VEGF-A (200 ng/pellet; d) or FGF-2 (100 ng/pellet; e) on day 7 after implanting Hydron pellets into mouse corneas (original magnification ×20). We performed quantitative analysis of neovascularization of each group on day 7. Areas are expressed in mm2 (n = 4 mice per group). We present data as means ± SE (c–e), and SD (b) of n observations, and we identified differences using two-tailed t-test, **P < 0.01.
