Anti c myc

WP1130 were purchased from Selleck Chemicals LLC. DMDA-pateamine A (D-PatA) was kindly made available to us by Dr Daniel Romo (Department of Chemistry, Texas A&M University, College Station, TX, USA). Sodium arsenite, cycloheximide, puromycin, IgG agarose, streptavidin-linked agarose and anti-FLAG M2 agarose were purchased from Sigma Aldrich. Anti-eIF4B monoclonal, anti-USP9X, and anti-eIF3c, anti-eIF4AI/II, GAPDH antibodies were purchased from Santa Cruz Biotechnology. Anti-c-Myc was from Abways Technology. Anti-XIAP antibody was obtained from BD Biosciences. Rabbit anti-human eIF4B polyclonal antibody was kindly provided by Dr John W.B. Hershey at UC Davis. Rabbit polyclonal Dcp1a antibody was kindly provided by Dr. Jens Lyke-Andersen at UC San Diego. Anti-Bcl-2 antibody was from Abcam Inc. anti-β-actin, β-tubulin and secondary antibodies conjugated with HRP were purchased from Kang-Cheng Biotech. Secondary antibodies conjugated with fluorescence were from Jackson ImmunoResearch Labs.

Total proteins were extracted from the cells and tissues using radio immunoprecipitation assay buffer (BIOSS) mixed with phenylmethylsulfonyl fluoride (BIOSS) and quantified using bicinchoninic acid assay (Beyotime Institute of Biotechnology). Protein extractions (30ug per well) were separated using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes (MilliporeSigma). The membranes were blocked with 5% fat-free dried milk for 1 h at room temperature. After incubation with high-affinity anti-FTO (1:1000, Abcam, USA), anti-STAT3 (1:1000, Cell Signaling Technology, USA), anti- phosphorylation-STAT3 (1:1000, Cell Signaling Technology, USA), anti-Bcl-2 (1:1000, Cell Signaling Technology, USA), anti-c-Myc (1:1000, Abways Technology, China), anti-CyclinD-1 (1:1000, Abways Technology, China), anti-β-actin (1:5 000, Bioss, China), or anti-GAPDH (1:5000, Bioss, China) antibodies at 4°C overnight, the membranes were incubated with the HRP-conjugated secondary antibodies goat anti-rabbit (1:10,000; Boster, China) or goat anti-mouse (1:5000; Boster, China) for 1 h at room temperature. Proteins were detected using BeyoECL chemiluminescence kit (Biyuntian, China) and detected using the Amersham ImageQuant 800 system (Cytiva). The density of bands were measured using ImageJ (v1.51, National Institutes of Health).