Plata-Gómez, Ana BelénCrespo, Maríade la Calle Arregui, Celiade Prado-Rivas, LucíaSabio, GuadalupeEfeyan, Alejo2024-03-112024-03-112021-12-17STAR Protoc. 2021 Oct 29;2(4):100918.http://hdl.handle.net/20.500.12105/18915We present a protocol for measuring the activity of the mechanistic target of rapamycin (mTOR) pathway in ex vivo isolated mouse primary hepatocytes. It can be used as a tool for genetic, pharmacological, metabolomic, and signal transduction procedures. We discuss critical aspects for improving yield, viability, and modulation of the mTOR pathway. This protocol can be adapted to other signaling cascades and is compatible with multiple readouts. For complete details on the use and execution of this protocol, please refer to Ortega-Molina et al. (2021).engVoRhttp://creativecommons.org/licenses/by-nc-nd/4.0/TOR Serine-Threonine KinasesAnimalsCell Culture TechniquesCells, CulturedHepatocytesLiverMiceSignal TransductionAttribution-NonCommercial-NoDerivatives 4.0 Internacional347551202410091810.1016/j.xpro.2021.1009182666-1667STAR protocolsopen access