Snu 407

Human HT-29 colon cancer and SNU-407 cells were obtained from the Korean Cell Line Bank (Seoul, Republic of Korea). Normal human FHC colon cells were obtained from the American Type Culture Collection (Rockville, MD, USA). HT-29 and SNU-407 cells were cultured in RPMI-1640 medium (Invitrogen, Grand Island, NY, USA) that contained 10% heat-inactivated fetal bovine serum (Sigma-Aldrich Co.). FHC cells were cultured in a 1:1 mixture of Ham’s F12 and DMEM that contained HEPES (25 mM), cholera toxin (10 ng/ml), insulin (5 μg/ml), transferrin (5 μg/ml), hydrocortisone (100 ng/ml), and 10% fetal bovine serum.

Six human colon cancer cell lines with documented Kras G12D mutations (Kras^G12D), LS-174T, SNU-407, and SNU-C2A, and three Kras wild-type (Kras^WT) colon cancer cells, Caco2, COLO-320DM, and HT29, were purchased from American Type Culture Collection (ATCC, Manassas, VA. USA) or Korea Cell Line Bank (KCLB, Seoul, Korea). Enriched cells were then cultured at sub-clonal density (1–10 cells/cm²) with complete growth medium of each cell line at 37 °C and 5% CO₂ atmosphere. Caco2 was cultured in MEM medium (Gibco-Invitrogen, Waltham, MA, USA) supplemented with 20% FBS (Gibco-Invitrogen) at 37 °C and 5% CO₂; LS-174T, SNU-407, SNU-C2A, HT-29, and COLO-320DM were cultured in RPMI 1640 medium (Gibco-Invitrogen) with 10% FBS. For the palmitate treatments, cells were treated with 50 μM conjugated palmitate (Sigma-Aldrich) with 10% (w/v) fatty acid free BSA (Sigma-Aldrich) in the DMEM medium. Cell lines used in silico analysis, in vitro, and in vivo experiments, as listed in Table S2.