A5256701

The HCT116, HT29, MDA-MB-231, SW480, CaCO2, HCC70, MCF7, AGS, and NCI-N87 cell lines were purchased from ATCC. The MDA-MB-231-SMYD3-KO and HCT116-SMYD3-KO cell lines were generated using the CRISPR/Cas9 technology. The U2OS cell line was kindly provided by Prof. Jeremy Stark.
The HCT116-OXA-R cell line was established from the HCT116 parental cell line by continuous exposure to oxaliplatin with stepwise increasing concentrations ranging from 1 μM to 10 μM over approximately 6 months. The CRC cell lines HCT116, HT29, SW480, and CaCO2, and the BC cell lines MDA-MB-231 and MCF7, and the U2OS DR-GFP cell line were cultured in DMEM high glucose without pyruvate (41965–039, Gibco) with 10% FBS (A5256701, Gibco) and 100 IU/ml penicillin–streptomycin (15140–122, Gibco). The BC cell line HCC70 and the GC cell lines AGS and NCI-N87 were cultured in RPMI high glucose without pyruvate (21875–034, Gibco) with 10% FBS (A5256701, Gibco) and 100 IU/ml penicillin–streptomycin (15140–122, Gibco). All cell lines were tested to be mycoplasma-free (117048; Minerva Biolabs) multiple times throughout the study. All cell cultures were maintained in a humidified incubator at 37 °C and 5% CO2.

The human pancreatic cancer cell line MIA PaCa-2 (ATCC#: CRL-1420™) was used. For both in vitro and in vivo experiments, cell cultures of passages 3 to 10 and a confluence of 70–90% were used. For in vitro experiments, cultured cells were supplied with high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM, 11965092; Gibco, ThermoFisher Scientific, Waltham, MA, USA) enhanced with 10% fetal bovine serum (FBS, A5256701; Gibco, ThermoFisher Scientific, Waltham, MA, USA) and 4 mM GlutaMax (35050079; Gibco, ThermoFisher Scientific, Waltham, MA, USA). For in vivo experiments, cells were cultured in Ham’s F12 medium (21127030; Gibco, ThermoFisher Scientific, Waltham, MA, USA) enhanced with 2 mM L-glutamine (A2916801; Gibco, ThermoFisher Scientific, Waltham, MA, USA) and 10% FBS (Gibco, ThermoFisher Scientific, Waltham, MA, USA). For cell detachment from flasks, TrpyLE (12605010; Gibco, ThermoFisher Scientific, Waltham, MA, USA) was used. For cell fixation, paraformaldehyde (PFA, 047392.9M; Sigma-Aldrich, St. Louis, MO, USA) was used. For cell washing, phosphate-buffered saline (PBS, J61196.AP, ThermoFisher Scientific, Waltham, MA, USA) was used. All cell incubations were at 37 °C with 5% CO₂.