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. 2010 Aug 2;12(11):1088–1101. doi: 10.1093/neuonc/noq079

Fig. 5.

Fig. 5.

In vivo angiogenesis assay. Treatments (48 hours): scrambled siRNA, survivin siRNA, 1 µM 4-HPR, and survivin siRNA + 1 µM 4-HPR. (A) In vivo angiogenesis assay using U251MG cells for implantation. The cells were treated for 48 hours, suspended in 200 µL of serum-free medium, injected into a diffusion chamber, implanted under the dorsal skin of nude mice, and left for 10 days. Strong development of tumor-induced neovasculature (TN) arising from pre-existing vessels (PV) as curved thin structures in zigzag pattern was observed in U251MG control (not shown) and scrambled siRNA transfected cells. The formation of such microvasculature was considerably reduced in both survivin siRNA- and 4-HPR-treated cells and almost completely inhibited after treatment with a combination of both agents. (B) Quantitative presentation of in vivo angiogenesis. The TN was measured with the help of an ocular micrometer. Values are mean ± SD of 6 samples from each group (*P < .001 compared with the control mean values and #P < .001 compared with the survivin siRNA or 4-HPR mean values). (C) A nude mouse bearing a diffusion chamber implanted under the dorsal skin for the in vivo angiogenesis assay.