Su11274

Cell lines (LOVO, HUVEC, CT26 and SW48) were obtained from the American Type Culture Collection. The SW48, LOVO, and HUVEC were incubated in Dulbecco’s modified Eagle’s medium (DMEM) with high glucose content (Invitrogen, CA, USA) containing 10% fetal bovine serum (FBS; gibco, USA) and 1% penicillin/streptomycin, CT26 was maintained in RPMI1640 (Invitrogen, CA, USA) supplemented with 10% FBS and 1% penicillin/streptomycin. All cells were cultured in humidified atmosphere of 5% CO2 in air at 37 °C. SU11274 (Medchem express, USA), a specific chemical inhibitor of Met; Anti-HGF antibody (Sinobiological, China), a neutralizing antibody for HGF; recombinant human HGF (Peprotech, USA). GFP lentiviral particles were purchased from Genechem Co (china, shanghai). The transfection of lentiviral was conducted following instruction of manufacturer. For lentiviral transduction, 5000 cells/well were seeded on 96 well tissue culture plates and infected the following day with lentiviral particles (Santa-Cruz Biotechnology, Dallas, TX) at a MOI of 10 in the presence of 10 mg/ml polybrene, purchased from). GFP-expressing cells were selected with 2 μg/ml puromycin (Medchem express, USA) and enriched by three cycles of fluorescence-activated cell sorting (FACS).

For GC cell treatment, we used the MET inhibitor SU11274 (HY-12014, MedChemExpress, Monmouth Junction, NJ, USA) and the SMYD3 inhibitor EM127, which was synthesized as previously described in Parenti et al. [14 (link)]. Both inhibitors were dissolved in DMSO and stored at −80 °C. For each inhibitor, concentrations and treatment duration are indicated in the figure legends.