Pf 04217903

For in vitro assays, NZ001 was obtained from Nanjing Zhongrunyuan Pharmaceutical Company (Nanjing, Jiangsu, China). XL184, sorafenib and PF-04217903 were purchased from Selleck Chemicals (Houston, TX, USA). Goat anti-mouse VEGF antibody (AF-493-NA) was purchased from R&D Systems (Minneapolis, MN, USA). NZ001, XL184 and PF-04217903 were prepared as a 20-mM stock solution in DMSO (Sigma-Aldrich, St. Louis, USA) for in vitro studies. For in vivo studies, NZ001 was formulated in sterile ddH2O and administered via oral gavage at 10 mg/kg or 30 mg/kg. sorafenib was dissolved in Cremophor EL/ethanol (50:50; Sigma Cremophor EL, 95% ethyl alcohol) at 4-fold (4×) the highest dose. The final dosing solutions were prepared on the day of use by diluting to 1× with ddH2O and were administered via oral gavage at 30 mg/kg. VEGF antibody was injected intraperitoneally 3 times per week at 7.5 mg/kg. Recombinant human HGF, mouse HGF and human VEGF were obtained from R&D Systems (Minneapolis, MN, USA). All information of primary antibodies used for Western blot and immunoprecipitation were shown on Additional file 1: Table S1. All secondary antibodies were purchased from Jackson ImmunoResearch (Philadelphia, PA, USA).

NCI-N87, HGC-27 and SGC-7901 human GC cells were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). KATOIII, SNU16, MKN-45 and MKN-28 GC cells were obtained from 3DBiopharm Biotech Co. Ltd. (Shanghai, China). Cells were cultured in Minimum Essential Medium (HGC-27) or Roswell Park Memorial Institute 1640 medium containing 10% fetal bovine serum (rest of the cells) and 1% penicillin-streptomycin at 37°C in a humidified atmosphere with 5% CO₂. AZD4547, AZD8931, erlotinib and PF04217903 were purchased from Selleck Chemicals (Houston, TX, USA). Cetuximab was provided by Bristol-Myers Squibb (New York City, NY, USA). Recombinant human HGF, EGF, NRG1, PDGF and IGF-1were purchased from PeproTech (Rocky Hill, NJ, USA). The compounds were dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, St Louis, MO, USA) and diluted with culture medium to the desired concentration, with a final DMSO concentration < 0.2% (v/v). DMSO was also added to the control cells in culture.