F12k medium

A gastric epithelial cell line (GES-1) and five GC cell lines (AGS, HGC27, MGC803, NCI-N87 and SNU-1) were obtained from the Type Culture Collection of the Chinese Academy of Science (Shanghai, China) and were validated by short tandem repeat DNA profiling. Cells were cultured in RPMI-1640 (BBI Life Sciences, Shanghai, China) or F12K medium (Zhong Qiao Xin Zhou Biotechnology, Shanghai, China) supplemented with 10% fetal bovine serum (FBS), 100 µg/ml penicillin, and 100 mg/ml streptomycin at 37°C with 5% CO₂ in a humidified incubator (Thermo Fisher Scientific, Inc., Waltham, MA, USA).

A gastric epithelial cell line (GES-1) and GC cell lines (AGS, HGC27, NCI-N87, MGC803 and SNU-1) were obtained from the Type Culture Collection of the Chinese Academy of Sciences. All of the cells were validated by short tandem repeat DNA profiling and were confirmed negative for Mycoplasma contamination. Cells were cultured in RPMI-1640 (BBI Life Sciences Corporation) or F12K medium (Zhong Qiao Xin Zhou Biotechnology Co., Ltd.) supplemented with 10% fetal bovine serum (cat. no. E600001; Sangon Biotech Co., Ltd.), 100 µg/ml penicillin, and 100 mg/ml streptomycin at 37°C with 5% CO₂ in a humidified incubator (Thermo Fisher Scientific, Inc.).
Construction of TAFA5-targeting short hairpin (sh)-RNAs and packaging of lentiviruses. Two targeting shRNAs (shTAFA5-1 and shTAFA5-2) and a nontargeting scrambled RNA (scramble) were subcloned into the GV248 lentivirus vector by Shanghai GeneChem Co., Ltd. The shTAFA5-1 target sequence was 5′-CGCAAGAATCATCAAGACCAA-3′, and the shTAFA5-2 target sequence was 5′-CACCTGTGAGATTGTGACCTT-3′. Lentiviral stocks were prepared, as previously described (15 (link)).