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Determination of Polycation-Mediated Perturbation of Mitochondrial Respiration in Intact Cells by High-Resolution Respirometry (Oxygraph-2k, OROBOROS)

Lookup NU author(s): Professor Moein MoghimiORCiD


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A better understanding of the molecular basis of polycation-mediated impairment of mitochondrial bioenergetics might improve the design and synthesis of more efficient and safer polymeric transfectants. Here we utilize the phosphorylation control protocol for studying the effect of polycations on mitochondrial respiration in intact mammalian cells using Oxygraph-2k (OROBOROS). The protocol offers an opportunity to comprehensively monitor mitochondrial respiration through consecutive additions of various cell membrane permeable compounds that alter mitochondrial respiration, thus providing useful information on different states of mitochondrial respiration. Furthermore, we demonstrate how to analyze the data obtained with the phosphorylation control protocol and how to calculate the respiratory flux ratios, which can be used as indicators of respiratory functionality and mitochondrial health.

Publication metadata

Author(s): Hall A, Moghimi SM

Publication type: Book Chapter

Publication status: Published

Book Title: Nanotechnology for Nucleic Acid Delivery: Methods and Protocols

Year: 2019

Volume: 1943

Pages: 313-322

Online publication date: 06/03/2019

Acceptance date: 02/04/2018

Series Title: Methods in Molecular Biology

Publisher: Humana Press

Place Published: New York


DOI: 10.1007/978-1-4939-9092-4_20

PubMed id: 30838625

Library holdings: Search Newcastle University Library for this item

ISBN: 9781493990917