Decitabine

These inhibitors (C646, GSK343 and decitabine; MedChemExpress, Shanghai, China) were initially dissolved in 10 mM DMSO to a range of concentrations (C646: 1, 2, 4, 6, 8, and 10 μM; GSK343: 1, 2, 4, 6, 8, and 10 μM; decitabine: 1, 2, 4, 6, 8, and 10 μM) with FGSC medium. The FGSCs were cultured to a density of 40% on a 48-well plate and treated with inhibitor for 24 h. Finally, the appropriate concentrations of different inhibitors (C646: 4 μM; GSK343: 4 μM; decitabine: 8 μM) were determined according to the results of CCK8.

Recombinant Human TNF-α (Cat. No. 300-01A), IFN-γ (Cat. No. 300-02), and insulin growth factor (IGF; Cat. No. 100-11) were purchased from PeproTech (Cranbury, NJ, USA). Sodium butyrate (Cat. No. S1200) and cisplatin (Cat. No. C2210000) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). The inhibitors CUCD-101 (Cat. No. HY-10223), decitabine (HY-A0004), santacruzmate A (HY-N0931), zebularine (HY-13420), olaparib (HY-10162), alpelisib (HY-15244), and MK-2206 (HY-108232) were all purchased from MedChemExpress (Sollentuna, Sweden). Fulvestrant (ICI 182,780; Cat. No. 1047) was obtained from Tocris Bioscience (Bristol, UK). A medium containing 0.5% DMSO was used as a vehicle-only control for the experiments.

The cell culture adapted Eμ-Myc lymphoma cell clone 152M was maintained in RPMI 1640 medium (Sigma-Aldrich, Taufkirchen, Germany) supplemented with 10% (v/v) fetal calf serum (FCS), 50 µM β-mercaptoethanol (Carl Roth, Darmstadt, Germany), 10 mM HEPES and 100 units/mL penicillin and 0.1 mg/mL streptomycin (both Sigma). Cells were cultured under humidified atmosphere at 5% CO₂ and 37 °C. For inhibition of DNA methyltransferases, cells were treated with 0.1, 0.5, and 1.0 µM Decitabine (MedChemExpress, Monomouth Junction, NJ, USA) for 48 h. Cell viability was monitored by Trypan Blue (Sigma-Aldrich, Taufkirchen, Germany) staining.