Netrin-4 induces HMVEC-dLy proliferation, migration, tube formation and survival. (A) Mitogenic potential of different doses of Netrin-4 on lymphatic dermal human microvascular endothelial cells (HMVEC-dLys) compared with several known lymphangiogenic growth factors (FGF-2 or bFGF, HGF, VEGF-A (VA), VEGF-C (VC), and complete (CM) or basal cell (BM) culture media. Cell number was assessed using dojindo reagent 72 hours after treatment and expressed as fold increase versus control. (B) HMVEC-dLy proliferation under a single dose of Netrin-4 (500 ng/mL), FGF-2 (40 ng/mL), VEGF-A (50 ng/mL), or VEGF-C (300 ng/mL) assessed every 24 hours for 3 days. (C) Chemotactic effects of different doses of Netrin-4 and VEGF-C on HMVEC-dLys in a Boyden chamber assay. (D) HMVEC-dLy adhesion on various matrixes: Fibronectin (FN), Netrin-4, Collagen I (Col. I), and Poly-L-Lysin (PLL) at 10 μg/mL. (E) In vitro tube formation by HMVEC-dLys under different doses of Netrin-4, FGF-2 (bF), HGF, VEGF-A (VA), VEGF-C (VC), or complete media (CM). (F) Inhibition of serum deprivation–induced HMVEC-dLys apoptosis under different doses of Netrin-4, FGF-2, VEGF-A (VA), VEGF-C (VC), and complete media (CM). *P < .05.