Het 1a cell

SV40 large T transfected esophageal epithelial HET 1-A cells (ATCC, Manassas, VA, USA) were used as a representative of benign esophageal mucosa. Human hTERT-immortalized nonneoplastic Bar-T cells were used as a representative of Barrett’s Esophagus mucosa. Bar-T cells were a generous gift from R. Souza and S.J. Spechler, Department of Medicine, VA North Texas Health Care System and the University of Texas Southwestern Medical School, Dallas, TX, USA. OE-19 cells (Sigma-Aldrich, St Louis, MO, USA), derived from an adenocarcinoma of a gastric/esophageal junction of a 72 year-old male patient, were used as a representative of esophageal adenocarcinoma. HET 1-A cells were maintained in Bronchial Epithelial Cell medium (BEGM BulletKit, Clonetics Corporation, Walkersville, MD, USA) supplemented with penicillin (100 U/ml), streptomycin (100 μg/ml) and 10% fetal bovine serum (FBS). Bar-T cells were maintained in keratinocyte basal medium 2 (KBM-2; Clonetics Walkersville, MD, USA) and OE-19 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen, Carlsbad, CA, USA), respectively, supplemented with penicillin (100/ml), streptomycin (100 μg/ml) and 10% FBS (Invitrogen, Carlsbad, CA, USA). The cells were cultured in 75 ml flasks (Greiner Bio-One, Monroe, NC, USA) at 37 °C with 5% CO₂. All cell lines have been authenticated (Bio-Synthesis Inc., Lewisville, TX, USA).

Human ESCC KYSE-30, KYSE-150, KYSE-270, and KYSE-410 cell lines were obtained from Dr. C.S. Yang at Rutgers University and they are cultured in RPMI-1640/Ham’s F12 (1:1) medium with 5% FBS^{53 (link)}; EAC cell line OE33 (Sigma-Aldrich, USA) was cultured in RPMI-1640 medium containing 10% FBS; normal esophageal epithelial HET-1A cells (ATCC, USA) were maintained in serum-free LHC-9 medium^{53 (link)}. ATAC-seq analysis has been conducted in HET-1A and ESCC cell lines. The transcriptome profiling is consistent to published data. Cells are periodically examined to ensure free from mycoplasma contamination using Commercial Detection Kit (Lonza, Switzerland, LT07-703). One week before experiment, HET-1A cells were adapted in experimental culture medium same as that used for ESCC cells. All culture medium contained 1% penicillin/streptomycin and cells were maintained at 37 °C in a humidified 5% CO₂ incubator.