All cell lines were grown at 37°C in 5% CO2. Colo205 cells were cultured in RPMI 1640 [10% fetal bovine serum (FBS), Hepes, pyruvate, 1% penicillin/streptomycin (P/S)]. HCT116 and HT29 were cultured in McCoy’s (10% FBS, 1% P/S). RKO cells were cultured in MEM Earle’s [10% FBS, pyruvate, NEAA (nonessential amino acid), 1% P/S]. LoVo cells were cultured in F-12K (10% FBS, 1% P/S). T84 cells were cultured in 50/50 mix of HAM’s F-12 and Dulbecco’s modified Eagle’s medium (DMEM) (2.5 mM l-glutamine, 5% FBS, 1% P/S). Colo320HSR, DLD-1, HCT15, and patient-derived cell lines CRC247, CRC12x, and CRC119 were cultured in RPMI 1640 (10% FBS, 1% P/S). All cell lines were purchased within 6 months from Duke Cell Culture Facility (CCF) or the American Type Culture Collection, and patient-derived lines were obtained from D. Hsu (Duke University) and passaged less than 10 times. All cell lines were authenticated using Promega PowerPlex 18D kit for short tandem repeat (STR) analysis or were purchased within 6 months from Duke CCF. All cell lines were tested for mycoplasma by Duke CCF. FBS was purchased from Gibco, and P/S was purchased from Thermo Fisher Scientific. Medium was purchased from Gibco or Sigma-Aldrich.

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