Figure 5.

Anti-VEGF antibody blocks GDF15 mediated angiogenesis. (A) Wound healing assay was performed using human brain microvascular endothelial cells (HBMVECs) in the absence/presence of anti-VEGF antibody. The day before the experiment, HBMVECs were seeded in 12-well plates (1.5 × 10⁵/well). The monolayer was scratched to introduce a wound and cells were cultured with control conditioned medium (Con-CM; conditioned medium from HBMVECs) or GDF15-CM HBMVECs (conditioned medium from rhGDF15-stimulated HBMVECs) with added control antibody or α-VEGF (1 ng/mL each). The wounded areas were photographed at 0 h and 24 h and the cell migration as percent mean distance migration was assessed. (B) Representative photographs of tube formation assay. HBMVECs (1.5 × 10⁵ cells) were incubated on Matrigel matrix for 12 h with Con-CM or GDF15-CM in the presence or absence of α-VEGF (1 ng/mL). The tube length was measured using ImageJ angiogenesis analyzer (significance was compared with the group Con-CM in the absence of α-VEGF). Data are presented as the mean ± standard error (*p < 0.01 and ****p < 0.001 compared with the non-treated CM control; ^# p < 0.05 and ^### p < 0.001 compared with rhGDF15- CM group).