The anti-CD146 mAb AA98 blocks VEGF-induced VEGFR-2 phosphorylation and Akt/p38/NF-κB activation. (A) Phosphorylation of VEGFR-2 induced by VEGF in HUVECs was assessed in the presence of anti-CD146 mAbs AA98 or AA1. Total VEGFR-2 quantification was determined by measuring the band density and then normalizing against internal controls. Results are presented as the means ± SEM of normalized values from 3 independent assays. (B) P38 activation induced by VEGF in the presence of AA98 or AA1. (C-D) NF-κB (p-p65 and IκBα) activation by VEGF in the presence of AA98 or AA1. (E) Akt activation induced by VEGF in the presence of AA98 or AA1. (F) Immunofluorescence showing inhibition of AA98 on the translocation of p65 and p50 from the cytoplasm to nucleus. NF-κB activation was detected using specific anti-p65 (red) and anti-p50 (red) Abs (scale bar indicates 20 μm). *P < .05; **P < .01.