Ptyr1175 vegfr 2

3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT), dimethyl sulfoxide (DMSO), M199 and Lectin from Bandeiraea simplicifolia (Griffonia simplicifolia) were purchased from Sigma Chemical Co. (St. Louis, MO). Recombinant human VEGF₁₆₅ and human basic fibroblast growth factor (bFGF), were obtained from PeproTech Inc. Recombinant human EGF was purchased from ProSpec Company. Matrigel were purchased from BD Bioscience (San Diego, CA). The primary antibodies for VEGF, VEGFR-2, p^Tyr1175-VEGFR-2, p^Tyr951-VEGFR-2, p-p38, p-ERK, p38, ERK, Akt, p-Akt, PI3K p110α, PI3K p85, p-PTEN, p-Colifin, Colifin, cleaved-caspase3/9, cleaved-PARP and all the secondary antibodies were acquired from Cell Signaling Technology (Cell Signaling Technology, Inc, USA). The primary antibodies for GAPDH, β-actin, Bax and Bcl-2 were obtained from Proteintech Group (Proteintech Group, Inc., USA). Annexin V-FITC apoptosis detection kit was purchased from Roche (Indianapolis, IN). Control siRNA (5′-uucuuccgaacgugucacgutt-3′) and siRNA against human VEGFR2 (5′-gcggcuaccaguccggauatt-3′) were synthesized by genepharm. Trypsin and DMEM/F12 were obtained from HyClone, and fetal bovine serum (FBS) was obtained from Gibico. M199 Medium was bought from LIFE. siRNA and all other chemicals were purchased from Sigma Chemical Co. unless otherwise specified.

Antibodies used in western blots are as follows: β-actin (RM2001, Ray Antibody Biotech, Beijing, China), α-tubulin (RM2007, Ray Antibody Biotech, Beijing, China), MIG6 (WH0054206M1, Sigma), pTyr1175-VEGFR2 (2478, Cell Signaling), pTyr951-VEGFR2 (2471, Cell Signaling), pTyr1054/59-VEGFR2 (44-1047G, Invitrogen), pTyr1214-VEGFR2 (AF1766, R&D Systems), VEGFR2 (9698, Cell Signaling), pTyr783-PLCγ1 (2821S, Cell Signaling), PLCγ1 (5690S, Cell Signaling), pSer1177-eNOS (9570, Cell Signaling), and eNOS (32027, Cell Signaling).