Protocol for detecting mitochondria extracellular vesicles of brown adipose tissue in mice

Flavia Tortolici; Claudia Di Biagio; Daniele Lettieri-Barbato; Katia Aquilano

STAR Protocols (Sep 2024)

Protocol for detecting mitochondria extracellular vesicles of brown adipose tissue in mice

Flavia Tortolici,
Claudia Di Biagio,
Daniele Lettieri-Barbato,
Katia Aquilano

Affiliations

Flavia Tortolici: University of Rome, “Tor Vergata,” Biology Department, 00133 Rome, Italy
Claudia Di Biagio: University of Rome, “Tor Vergata,” Biology Department, 00133 Rome, Italy
Daniele Lettieri-Barbato: University of Rome, “Tor Vergata,” Biology Department, 00133 Rome, Italy; Corresponding author
Katia Aquilano: University of Rome, “Tor Vergata,” Biology Department, 00133 Rome, Italy; Corresponding author

Journal volume & issue: Vol. 5, no. 3
p. 103161

Abstract

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Summary: Brown adipose tissue (BAT) is mitochondria rich, enabling high oxidative metabolism for non-shivering thermogenesis. The release of large/small extracellular vesicles (EVs) containing mitochondria or mitochondrial fragments, termed mito-EVs, may support mitochondrial quality control or intercellular communication. We present a protocol to isolate and characterize mito-EVs. We detail steps for BAT processing, cell debris removal, differential centrifugation (dC), and mito-EV analysis by flow cytometry and immunoblotting assays.For complete details on the use and execution of this protocol, please refer to Rosina et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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