Measuring Cellular Bioenergetic Parameters

ROS production was assessed using a fluorogenic probe (CellRox Deep Red Reagent, Thermo Fisher Scientific) as described previously [24 (link)]. Adenosine triphosphate (ATP) concentration was assessed using the Colorimetric ATP Assay Kit (Abcam, Cambridge, UK). ATP generation was normalized by the number of cells. The GSH concentration was calculated using the GSH-Glo Assay kit (Promega). Cellular NADPH contents were determined by a colorimetric determination method using an NADP/NADPH determination kit (Biovision, Milpitas, CA, USA). Mitochondrial membrane potential was measured by using the JC-1 Mitochondrial Membrane Potential Assay Kit (Cayman Chemical, Ann Arbor, MI, USA). All procedures were conducted according to the manufacturer's instructions.