4.6. Cell Proliferation Assay

CellTiter-Glo Luminescent Cell Viability Assay (Promega, G7572, Madison, WI, USA) was used to determine the number of viable cells in culture. Briefly, 5 × 10³ cell/well were seeded in 96-well plates in triplicate. After transfection of miRNAs for 3 days, a volume of CellTiter-Glo reagent equal to the volume of cell culture medium was added to each well. The plates were shaken for 2 min to induce cell lysis and further incubated for 10 min to stabilize the luminescent signal. As there is a linear relationship between the luminescent signal and the number of cells, cell proliferation was measured by reading the absorbance at 490 nm using a GloMax 96 Microplate Luminometer (Promega, Madison, WI, USA). Fold change was calculated as the ratio of recorded luminescence values.