Spectra gemini spectrofluorimeter

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.

MLTC-1 cells were seeded in 96-well plates at 100,000 cells/well. Two days later, the medium was replaced with serum-free medium in the absence (control) or presence of FLX (0–100μM), MET (0–1000μM), or A-769662 (0–1000μM) for 1 hour at 37°C before the addition of 20μl of CellTiter-Blue Reagent (Promega, Madison, WI, USA) to each well. After having incubated for 2 hours at 37°C, changes in fluorescence were recorded with a Spectra Gemini spectrofluorimeter (Molecular Devices, Sunnyvale, CA) at an excitation wavelength of 560nm and an emission wavelength of 590nm. The fluorescent signal from the CellTiter-Blue Reagent is proportional to the number of viable cells.