The following antibodies were used in this study: anti-CD146 rabbit polyclonal antibody and anti-CD146 mouse mAb of AA1 were generated in our lab. Anti-β-actin antibody, HRP-conjugated goat anti-mouse and anti-rabbit IgG antibodies were from Sigma-Aldrich. Anti-human IgG Fc and anti-His antibodies were from ZSGB-BIO. Anti-VEGFR-3 antibody and anti-VEGF-C antibody were from Abcam. Anti-AKT, anti-p-AKT, anti-p38, anti-p-p38, anti-ERK and anti-p-ERK antibodies were from Cell Signaling Technology. Goat anti-mouse Alexa Fluor 488 and goat anti-rabbit Alexa Fluor 555 were from Invitrogen.
The following reagents were used: recombinant human VEGFR-3-Fc, human VEGF-C, Fc-CD146 and sCD146 were from Sino Biological. Human VEGF-C156S was from R&D. ERK1/2 inhibitor, SCH772984, and p38 inhibitor, FHPI, were from Selleckchem. Growth factor-reduced Matrigel was from BD Biosciences. Fugene HD, DAPI and protease inhibitor cocktails were from Roche. Protein G sepharose beads was from Santa Cruz. Enhanced Chemiluminescence Assay Kit for WB was from Pierce. Cell Counting Kit-8 (CCK-8) for cell proliferation assay was from Dojindo. Human Fc, Carboxymethylcellulose for spheroid assay and Fluorescein isothiocyanate-dextran average molecular weight 2000 KD and Tetramethylrhodamine-dextran average molecular weight 70 KD were from Sigma-Aldrich.
The following reagents were used: recombinant human VEGFR-3-Fc, human VEGF-C, Fc-CD146 and sCD146 were from Sino Biological. Human VEGF-C156S was from R&D. ERK1/2 inhibitor, SCH772984, and p38 inhibitor, FHPI, were from Selleckchem. Growth factor-reduced Matrigel was from BD Biosciences. Fugene HD, DAPI and protease inhibitor cocktails were from Roche. Protein G sepharose beads was from Santa Cruz. Enhanced Chemiluminescence Assay Kit for WB was from Pierce. Cell Counting Kit-8 (CCK-8) for cell proliferation assay was from Dojindo. Human Fc, Carboxymethylcellulose for spheroid assay and Fluorescein isothiocyanate-dextran average molecular weight 2000 KD and Tetramethylrhodamine-dextran average molecular weight 70 KD were from Sigma-Aldrich.