Ec109

Human ESCC cell lines (EC109, EC9706, TE1, KYSE30, KYSE140, KYSE150, KYSE410, KYSE450, and KYSE510) were obtained from Hunan Fenghui Biotechnology Co., Ltd. (Changsha, China) or Shanghai Fuxiang Biological Technology Co., Ltd. (Shanghai, China) and cultured in DMEM (EC109, TE1, and KYSE30 cells) or RPMI 1640 (EC9706, KYSE140, KYSE150, KYSE410, KYSE450, and KYSE510 cells) (Gibco, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, NY, USA) and penicillin-streptomycin solution (100 units/ml) (Solarbio, Beijing, China). WNT5A, HDAC7, and empty vector lentiviruses were obtained from GeneChem (Shanghai, China), and their detailed information is listed in Supplementary Tables 3, 4. EC109, EC9706, KYSE30, and KYSE450 cells were infected with lentiviruses according to the protocol provided by GeneChem Corporation.

Human ESCC cell lines (EC109, EC9706, KYSE150 and TE-1) and HEK-293T were obtained from the Hunan Fenghui Biotechnology Co., Ltd. (Changsha, China). These cells were cultured in high glucose DMEM medium (C11995500, Gibco, NY, USA) supplemented with 10% fetal bovine serum (04-001-1A, FBS, Biological Industries, Israel) and penicillin–streptomycin solution (100 U/ml, P1400, Solarbio, Beijing, China). Melatonin (#14427, Cayman Chemical Company, Michigan, USA), SAHA (HY-10221, MedChemExpress, USA), Lf3 (HY-101486, MedChemExpress, USA) and 10058-F4 (HY-12702, MedChemExpress, USA) stock solution was prepared in DMSO and diluted in culture media immediately prior to the subsequent experiments. The ESCC cells were treated with these chemical compounds for 48 h and then harvested for further analyses.