Colo205

The human colon cancer cell lines COLO-320, COLO205 and Caco-2 were purchased from the RIKEN Bioresource Center (Ibaraki, Japan). These cell lines were maintained at 37°C in RPMI 1640 medium (COLO-320 and COLO205) or MEM (Caco-2) containing 10% foetal calf serum (FCS) in a humidified atmosphere with 5% CO₂. To establish 5-FU-resistant Caco-2 cells, Caco-2 cells were maintained in MEM containing 10% FCS with continuous exposure to 5-FU at the concentration of 2 μM for 12 weeks.

Five human colorectal cancer cell lines (HCT116, HT29, SW480, Caco-2, and Colo205) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan). Dulbecco’s Modified Eagle’s Medium containing 10% foetal bovine serum, 100 U/ml penicillin and 0.1 mg/ml streptomycin served as the cell culture medium throughout all experiments. The cells were maintained at 37 °C in a humidified atmosphere of 5% CO₂ in air and passaged every 2 days to obtain exponential growth. All five cell lines were maintained in culture no more than 15 passages. All culture reagents were sourced from Gibco (Invitrogen, Carlsbad, CA, USA).

Human colon cancer cell lines HCT116, SW480, SW620, Caco2, HT29, Colo205 were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). Cells were maintained with Dulbecco's modified Eagle's medium (Thermo scientific, Barrington, IL) containing 10% fetal bovine serum (GIBCO, Grand Island, NY), 100 IU/ml penicillin, and 100 mg/ml streptomycin in a humidified tissue culture incubator at 37°C, 5% CO2. For C1GALT1 overexpression, HCT116 and SW480 cells were transfected with C1GALT1/pcDNA3.1/mycHis plasmids using Lipofectamine 2000 (Invitrogen, Life Technologies Inc., Grand Island, NY) according to the manufacturer's protocol. Empty pcDNA3.1/mycHis plasmids were used as mock transfection. Stable clones were selected with 400 μg/ml of G418 for 14 days. For C1GALT1 knockdown, HCT116 and SW620 cells were transfected with short hairpin (sh) RNA and selected with 2 μg/ml of puromycin for 14 days. The pLKO/C1GALT1-shRNA plasmid and non-targeting pLKO plasmids were purchased from National RNAi Core Facility (Academia Sinica, Taipei, Taiwan). The overexpression and knockdown of C1GALT1 were confirmed by western blotting.