Eso26

Three immortalized cell lines originated from Barrett’s esophagus, BAR10T (kindly provided by Dr. Rhonda Souza), CPA and CPB (ATCC, Manassas, VA, USA), were cultured with DMEM/F12 (GIBCO, New York, NY, USA), supplemented with 5% fetal bovine serum (FBS, GIBCO), 1% penicillin/streptomycin (GIBCO), 0.4 μg/ml hydrocortisone (Sigma-Aldrich, Saint Louis, MO, USA), 20 mg/L adenine hydrochloride hydrate (Sigma-Aldrich), 140 μg/ml bovine pituitary extract (Thermo Fisher Scientific, Waltham, MA, USA), Insulin-transferrin-sodium selenite media supplement (Sigma-Aldrich) and 20 ng/ml recombinant epidermal growth factor (Sigma-Aldrich). EAC cell lines, FLO1 (ATCC), OE19 (Sigma-Aldrich), SK-GT4 (kindly provided by Dr. Xiaochun Xu at MD Anderson), ESO26 (Sigma-Aldrich) and OE33 (kindly provided by Dr. David Beer) were cultured in DMEM or RPMI 1640 medium (GIBCO), supplemented with 10% FBS and 1% penicillin/streptomycin. All cell lines were grown at 37 °C in 5% CO₂. Cell lines were authenticated by Genetica DNA Laboratories using short tandem repeat profiling (Genetica DNA Laboratories, Burlington, NC, USA). Cells used were from the stocks immediately after authentication and cultured less than 6 months. Mycoplasma was tested periodically using the qRT-PCR method (Southernbiotech, Birmingham, AL, USA).

Human esophageal cancer cell lines (OE19, OE33, ESO26, KYSE270, SK-GT-2, Flo-1, ESO51, OE21, and OACM5.1C) were obtained from Sigma Aldrich (St. Lois, MO) and cultured according to manufacturer’s instructions. Flavopiridol was obtained from Cayman Chemical (Ann Arbor, MI), nanoparticle albumin-bound paclitaxel from Goshen Center for Cancer Care (Goshen, IN), BMS-754807 from Active Biochemical Limited (Maplewood, NJ), the cell proliferation reagent WST-1 from Roche Diagnostic Corporation (Indianapolis, IN), c-Myc siRNA (sc-29226) and control siRNA (sc-37007) from Santa Cruz Biotechnology (Santa Cruz, CA). pcDNA3-cMyc was a gift from Wafik El-Deiry (Addgene plasmid #16011) (Ricci et al., 2004 (link)) and pcDNA3-EGFP as a negative control (Addgene plasmid #13031) was purchased.