1. FAS1 domain protein inhibits association of αvβ3 integrin with VEGFR-2 and attenuates VEGF165-induced VEGFR-2 phosphorylation.
FAS1 domain protein inhibits association of αvβ3 integrin with VEGFR-2 and attenuates VEGF165-induced VEGFR-2 phosphorylation. A, serum-starved VEGFR-2/β3/HEK293 cells were plated on collagen-coated dishes in incomplete medium supplemented with the FAS1 domain protein (10 μmol/L) for 30 minutes and then stimulated for 5 minutes with VEGF165 (20 ng/mL) or vehicle alone. The cells were lysed and immunoprecipitated with anti-VEGFR-2 antibody. The immunoprecipitates were analyzed via SDS-PAGE followed by immunoblotting with anti-β3 integrin and anti-VEGFR-2 antibodies. The total cell lysates were immunoblotted with anti-β3 integrin antibody to assess its expression level. Immunoreactive bands were detected by ECL. B–E, serum-starved HUVEC, control shRNA/HUVEC, β3 integrin shRNA/HUVEC, VEGFR-2/β3/HEK293, and VEGFR-2/β5/HEK293 cells were plated on collagen-coated dishes in incomplete medium supplemented with the FAS1 domain protein (10 μmol/L) for 30 minutes and then stimulated for 5 minutes with VEGF165 (20 ng/mL) or vehicle alone. Cells were lysed and immunoprecipitated with anti-VEGFR-2 Ab. Immunoprecipitates were analyzed via SDS-PAGE followed by immunoblotting with anti-phosphotyrosine Ab and anti-VEGFR-2 Ab. The total cells were lysed and immunoblotted with anti-β3, anti-β5 integrin, anti-phosphotyrosine-VEGFR-2 Ab, and anti-β-actin Ab. Immunoreactive bands were detected by ECL. P, phosphorylated protein; IB, immunoblotting; IP, immunoprecipitation.
Ju-Ock Nam et al. Mol Cancer Res 2012;10:
©2012 by American Association for Cancer Research