Cell Viability Assay for MLTC-1 Cells

Cell viability was determined by CellTiter-Blue Reagent (Promega, Madison, WI, USA) using Spectra Gemini spectrofluorimeter (Molecular Devices, Sunnyvale, CA, USA) at an excitation wavelength of 560 nm and an emission wavelength of 590 nm according to the method described in our previous work [30 (link)]. MLTC-1 cells were cultured in 96-well plates with 100,000 cells/well at 37 °C under 5% CO₂ during two days. Then, the medium was removed and replaced with serum-free medium in the absence (control) or in the presence of KH7, 2-CE, 4-CE, LRE1, ddAdo3′, or ddAdo5′. Afterwards, the samples were incubated for 1 h at 37 °C before adding 20 µL of Reagent to each well and incubating it for 2 more hours at 37 °C. The fluorescent signal from the CellTiter-Blue Reagent is proportional to the number of viable cells.