Protocol for the assessment of mTOR activity in mouse primary hepatocytes

被引：4
作者：
Plata-Gomez, Ana Belen [1 ]
Crespo, Maria [2 ]
Arregui, Celia de la Calle [1 ]
de Prado-Rivas, Lucia [1 ]
Sabio, Guadalupe [2 ]
Efeyan, Alejo [1 ]
机构：
[1] Spanish Natl Canc Res Ctr CNIO, Metab & Cell Signaling Lab, Melchor Fernandez Almagro 3, Madrid 28029, Spain
[2] Ctr Nacl Invest Cardiovasc CN, Myocardial Pathophysiol, Melchor Fernandez Almagro 3, Madrid 28029, Spain
来源：
STAR PROTOCOLS | 2021年 / 2卷 / 04期
基金：
欧盟地平线“2020”;
关键词：
Cell culture; Cell isolation; Cell-based Assays; Metabolism; Metabolomics; Signal Transduction;
D O I：
10.1016/j.xpro.2021.100918
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
We present a protocol for measuring the activity of the mechanistic target of ra-pamycin (mTOR) pathway in ex vivo isolated mouse primary hepatocytes. It can be used as a tool for genetic, pharmacological, metabolomic, and signal trans-duction 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).
引用
收藏
页数：20
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