CellTiter-Glo Luminescent Cell Viability Assay

Cell viability was measured by determining the amount of adenosine triphosphate (ATP) synthesized inside the cells using the Promega CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega, Madison, WI). Total cell material was lysed, and ATP was collected on an opaque Corning 96-well plate (Corning, NY) using the reagents from the kit (CellTiter-Glo® Substrate (lyophilized), and CellTiter-Glo® Buffer) and the cell growth media associated with the designated cell type being analyzed. Luminescence was then detected using a SpectraMax® i3 Multi-Mode Microplate Detection Platform plate reader (Molecular Devices, San Jose, CA). An ATP standard curve was used as a positive control and as a quality control parameter to quantify the amount of ATP present based on the luminescence values. This control was made by combining stock ATP standard (Sigma-Aldrich, St. Louis, MO) with the working reagent and performing a serial dilution at μM concentrations of 50, 25, 12.5, 6.25, 3.125, and 0. Data shown are mean ± SEM from 3 separate biological replicates. A one-way ANOVA and Fisher’s LSD post-hoc test was used to determine statistically meaningful differences compared to the sham control exposed cells (p≤0.05). Statistical analysis was conducted using GraphPad Prism software (San Diego, CA).