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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Nat Cell Biol. 2017 Jan 23;19(2):120–132. doi: 10.1038/ncb3465

Figure 4.

Figure 4

Hypoxia does not significantly influence intravasation and extravasation. (a) Illustration of the CAM system. The tumour cells are seeded on the chorionic epithelium and penetrate this monolayer to reach the mesodermal vasculature. The CAM perpendicular opposite the implantation site, the lower CAM, is harvested 48 h after seeding. Following digestion of the CAM, the number of intravasated cells is counted. (b) Lower CAM cell suspension with MDA-231-HIF reporter cells pretreated for 72 h with 21% O2 (left) or 1% O2 (right). Scale bars, 25 μm; n = 3 CAMs per treatment were assessed; representative images are shown. (c) Quantification of cells counted in lower CAM. T-HEp3 GFP cells are used as a positive control. n = 3 CAMs with T-HEp3 GFP, n = 7 CAMs with MDA-MB-231-HIF reporter cells; bars indicate mean; Mann–Whitney test. (d) Post-hypoxia extravasation test. T-HEp3-GFP cells (3 × 105) were cultured in normoxia (21% O2) or hypoxia (1% O2) for 72 h and then tail vein injected in nude mice. Twenty-four hours later, the lungs were retrieved, collagenased and GFP+ cells were counted. n= 7 mice (normoxia), n= 6 mice (hypoxia); bars indicate mean; Mann–Whitney test.