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. 2015 Oct 6;5:14780. doi: 10.1038/srep14780

Figure 3. Atox1 promotes angiogenesis via activating Cu enzyme lysyl oxidase in ECs in an ATP7A-dependent manner.

Figure 3

(A) Sponge implant assay was performed by implanting polyvinyl alcohol sponge containing VEGF subcutaneously into WT and Atox1 KO mice. Representative images for H&E staining and isolectin immunostaining for blood vessel formation in sponges harvested on day 21. Right panels show quantitative analysis of the number of red blood cells (RBC) and isolection+ ECs. Scale bars = 50μm. (B) HUVECs were transfected with control, Atox1 or ATP7A siRNAs or treated with Cu chelator TTM (20 nM, 24 hrs) and seeded on Matrigel-coated plates in culture media containing VEGF for 6 h. Four random fields per well were imaged, and representative pictures are shown (left). Averaged numbers of capillary tube branches, branching points, and tube length per field are shown (Right). (C) HUVECs were treated with LOX inhibitor β-aminopropionitrile (BAPN, 100 μM) for 24 hrs and capillary tube formation was measured (n = 3). (D) Activity of LOX was measured in ischemic gastrocnemius muscle of WT and Atox1 KO mice (left)(n = 4) or in culture medium from VEGF (20ng/ml)-stimulated HUVECs transfected with siControl or siAtox1 (right)(n = 4). *p < 0.05.