On the day before treatments, cells were harvested, counted, seeded onto 96-well plates at a density of 104 cells per well, and adhered overnight.
For drug treatment, MRS-1220 (Tocris Bioscience, United Kingdom; CAS#183721-15-5) and CGS-15943 (Cayman Chemical Company, USA; CAS#104615-18-1) were prepared in a stock solution of respectively 10mM and 1.7mM DMSO (Merck KGaA, Germany). Serial dilutions using DMEM were performed to achieve a range of concentrations from 0.1nM to 100μM of the drugs. Control wells were treated with an equivalent volume of DMSO used for drug dilution. The treated and control cells were incubated for 72 h at 37°C, 5% CO2 incubator. All conditions were performed in triplicate to ensure reproducibility.
For drug treatment, MRS-1220 (Tocris Bioscience, United Kingdom; CAS#183721-15-5) and CGS-15943 (Cayman Chemical Company, USA; CAS#104615-18-1) were prepared in a stock solution of respectively 10mM and 1.7mM DMSO (Merck KGaA, Germany). Serial dilutions using DMEM were performed to achieve a range of concentrations from 0.1nM to 100μM of the drugs. Control wells were treated with an equivalent volume of DMSO used for drug dilution. The treated and control cells were incubated for 72 h at 37°C, 5% CO2 incubator. All conditions were performed in triplicate to ensure reproducibility.