Figure 3
From: CD146 is required for VEGF-C-induced lymphatic sprouting during lymphangiogenesis
CD146 directly binds to VEGF-C. (a) Co-immunoprecipitation assays in HDLECs. Cells were incubated with normal culture medium or VEGF-C156S (100 ng/ml) conditional medium. The cell lysates were prepared for immunoprecipitation with control mIgG or anti-CD146 mAb AA1. (b,d) HEK293 cells transfected with control empty or CD146 expression vectors were incubated with VEGF-C conditional medium or transfected with CD146 encoding vector without VEGF-C incubation. Binding of VEGF-C-His to cells was detected by co-immunoprecipitation (b) and immunofluorescence (d). Scale bars: 100 μm. (c) Direct in vitro interaction between VEGF-C and CD146 proteins. Purified Fc, Fc-CD146 or Fc-VEGFR-3 (200 ng/ml) was incubated with VEGF-C protein (200 ng/ml). The results were analyzed by WB. (e,f) defining the domain of CD146 required for interaction with VEGF-C. CD146 and its truncation mutants were expressed as Flag-tagged proteins. Immunoprecipitation was performed using anti-VEGF-C mAb. Full-length blots are presented in Supplementary Fig. S4.
