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In-vitro tube formation assay To determine the formation of tubes in vitro , 70,000 cells were seeded per 25 cm 2 flask and pre-treated with PBS, IGF-1 (100 nM) and cGP + IGF-1 (100 nM) in 0.2% serum media for 6 days before the cells were transferred onto growth factor-reduced Matrigel (GFR-Matrigel, BD, Franklin Lakes, NJ, USA)