2.9. Mitochondrial ATP synthase inhibition assay

Following miRNA transfection on day three, H1299 cells were treated with oligomycin A for 30 min. Since then the cellular ATP changes were measured by the following procedure with the CellTiter-Glo® Luminescent Cell Viability Assay (Promega) according to the manufacturer's instructions. We seeded the cells into 96 well plates at 5000 cells/well in 200 μL of media/well. After treatment of the compound, we incubated the plates at 37 with 5% CO2 for 24 h. We added a volume of CellTiter-Glo® 2.0 Reagent equal to the volume of cell culture medium present in each well, mixed the contents for 2 min on an orbital shaker to induce cell lysis to stabilize the luminescent signal, and incubated the plates for 10 min at room temperature. We read the luminescence by a GloMax® Discover Microplate Reader (Promega).