Snu 283

Human CRC cell lines (colo-201, colo-205, colo-320, HCT-15, HCT-116, HT-29, Lovo, LS174T, SNU-61, SNU-70, SNU-81, SNU-175, SNU-254, SNU-283, SNU-407, SNU-977, SNU-1033, SNU-1181, SNU-1235, SNU-1411, SNU-1544, SNU-1684, SNU-C1, SNU-C2A, SNU-C4, SNU-C5, SW-403, and SW-480) were purchased from the Korean Cell Line Bank [18 (link)]. All cell lines were cultured in RPMI1640 (Hyclone Laboratories, Ind., Logan, UT), supplemented with 10% fetal bovine serum, at 37°C in humidified 5% CO₂.

The NIH3T3 cell line stably expressing GLI-luciferase reporter construct was purchased from BPS Bioscience Inc., and maintained in DMEM (GenDEPOT, LLC) plus 10% fetal calf serum (Gibco; Thermo Fischer Scientific, Inc.) and 1% penicillin/streptomycin. Human CRC cell lines HCT116, SNU283, HCT8 (colon; colorectal) and HT29, DLD-1 (colon) were purchased from Korea Cell Line Bank (Seoul, Korea) and analyzed by STR profiling. HCT116 p53 knockout cell line was provided from the Keck School of Medicine of USC (Professor LIN ZHANG). Cells were cultured in RPMI-1640 medium (GenDEPOT, LLC) with 10% fetal bovine serum (Gibco; Thermo Fischer Scientific, Inc.), 1 mM L-glutamine, 26 mM sodium bicarbonate and 1% P/S (penicillin/streptomycin). All cells were cultured at 37°C in a 5% CO₂ incubator. All cell lines were confirmed not to be infected with mycoplasma using a mycoplasma detection kit (cat. no. SMD0173; BIOMAX).

The CRC cell lines used in this study were selected on the basis of DUSP4 expression data from the Cancer Cell Line Encyclopedia (https://portals.broadinstitute.org/ccle). Five CRC cell lines, COLO‐320, SNU‐283, CW‐2, SNU‐503, and SNU‐1033, express low levels of DUSP4, and three cell lines, SNU‐C2A, SNU‐C5, and HCT 116, express high levels of DUSP4. The CRC cell lines COLO‐320, CW‐2, and HCT 116 were obtained from RIKEN BioResource Center (Ibaraki, Japan), and SNU‐283, SNU‐503, SNU‐1033, SNU‐C2A, and SNU‐C5 were obtained from the Korean Cell Line Bank (Seoul, Korea). The human colonic epithelial cells (HCoEpiC) were purchased from ScienCell (Carlsbad, CA, USA). The human glioblastoma cell line A‐172, in which the promoter region of DUSP4 is known to be hypermethylated,9 was obtained from the Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku University (Miyagi, Japan). All of the cell lines were cultured in accordance with the suppliers' instructions. The genotypes of EGFR, KRAS, HRAS, NRAS, BRAF, APC, CTNNB1, TP53 and CDKN2A genes in all eight CRC cell lines were obtained from the Cancer Cell Line Encyclopedia. All of them except for HRAS, NRAS, and CDKN2A are summarized in Table 1, because these three genes are not mutated in any of the cell lines.