Fig. 2

Netrin-1 induces angiogenesis in vitro in HUVEC. a Netrin-1 (NTN-1) influence on HUVEC cell migration was determined using scratch assay, where cells were serum-starved and treated for 8 h with different recombinant human NTN-1 concentrations, as indicated. Endothelial basal media (EBM) was used as internal reference and recombinant human vascular endothelial growth factor (VEGF) (40 ng/mL) was used as a positive control. Representative pictures of each condition are shown (amplification × 10). b Quantified results correspond to the mean ± S.E.M. (n = 3, ^* p < 0.05). c The effect of recombinant human NTN-1 was determined in vitro to evaluate angiogenesis through tubule formation assay. HUVEC were serum-starved and seeded on Matrigel and treated with increased concentrations of NTN-1 for 4 h. EBM alone was used as internal reference and endothelial growth media (EGM) was used as a positive control. Representative images are shown for each experimental condition (scale bar = 15 μm). d Quantified results correspond to the mean ± S.E.M. (n = 6, ^* p < 0.05)