Abstract
Metabolic flexibility is vital for organisms to respond to and survive changes in energy availability. A critical metabolic flexibility regulator is peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), which regulates various transcription factors and nuclear receptors that, in turn, regulate mitochondrial homeostasis and fatty acid oxidation. PGC-1α is itself regulated, with one of the significant modes of regulation being acetylation. Thus, measuring the acetylation status of PGC-1α is a critical indicator of cells’ metabolic flexibility. In this chapter, we describe a method of evaluating PGC-1α acetylation in primary mouse myotubes. This method can also be used with other cell types and tissues.
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Acknowledgments
We would like to thank Eric L. Bell and Ana P. Gomes for help on advice and optimization on the immunoprecipitation and acetylation detection protocol. This work was supported by a fellowship from the Portuguese Foundation for Science and Technology (PD/BD/114173/2016) to J.A.A., the Paul F. Glenn Foundation for Medical Research, and NIA/NIH grant (R01DK100263) to D.A.S. Fig. 1 was created with BioRender.com.
Conflict of Interest: D.A.S is a consultant to, inventor of patents licensed to, and in some cases board member and investor of MetroBiotech, Cohbar, Life Biosciences and affiliates, InsideTracker, Zymo, EdenRoc Sciences and affiliates, Immetas, Segterra, Galilei Biosciences, and Iduna Therapeutics. He is also an inventor on patent applications licensed to Bayer Crops, Merck KGaA, and Elysium Health. Additional information https://u.nu/sinclair.
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Amorim, J.A., Sinclair, D.A. (2021). Measuring PGC-1α and Its Acetylation Status in Mouse Primary Myotubes. In: Palmeira, C.M., Rolo, A.P. (eds) Mitochondrial Regulation. Methods in Molecular Biology, vol 2310. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1433-4_17
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DOI: https://doi.org/10.1007/978-1-0716-1433-4_17
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