CRC Cell Lines and Drug Treatments

Human CRC cell lines, including HCT116, HT29, RKO, SW48, Lim1215, and NCI-H508, were purchased from the American Type Culture Collection and cultured in McCoy’s 5A modified media (Invitrogen). SNU-407 was purchased from AddexBio and cultured in RPMI 1640 (Invitrogen). DR5-knockout (KO) HCT116 cells were previously described (26 (link)). DR5-KO RKO cells were generated by CRISPR/Cas9 using a single guide RNA sequence 5’-CGCGGCGACAACGAGCACAA-3’ as described (27 (link)). Cells were authenticated in 2018 by genotyping and analysis of protein expression by western blotting, and routinely checked for Mycoplasma contamination by PCR. All cell lines were maintained at 37°C and 5% CO₂ atmosphere. Cell culture media were supplemented with 10% defined FBS (HyClone), 100 units/ml penicillin, and 100 μg/ml streptomycin (Invitrogen). For drug treatment, cells were plated in 12-well plates at 20–30% density 24 hr before treatment. DMSO (Sigma) stocks of JQ1, I-BET151 (Apexbio), OTX015, I-BET762, 5-FU, and oxaliplatin (Sigma) were prepared and diluted in cell culture media before adding to cells.