Rabbit anti hexokinase 2

Five gastric cancer cell lines (AGS, BGC-823, MKN-45, MGC-803, and SGC-7901) and one normal gastric mucosa endothelial cell line GES-1 were purchased from the Cell Research Institute of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI 1640 medium (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) and 100 units/ml penicillin plus 100 μg/ml streptomycin (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) at 37°C in a humidified cell culture incubator with 5% CO₂. The 5-Fu-resistant gastric cancer cell line AGS/5-Fu R was established according to the previous description by continually exposing to increased concentrations of 5-Fu. The acquired 5-Fu-resistant cells were re-selected by treating with 5-Fu at 300 uM every 2 months. The rabbit anti-GLUT1 (#73015), rabbit anti-Hexokinase 2 (#2867), rabbit anti-LDHA (#3582), and rabbit anti-β-actin (#4970) antibodies were purchased from the Cell Signaling Technology (Danvers, MA, USA). Mouse anti-PTBP1 (#32-4800) monoclonal antibody was purchased from Thermo Fisher Scientific (Shanghai, China). 5-Fu, 2-DG, and Oxamate were purchased from Sigma-Aldrich (Shanghai, China).