Mito-Kaede photoactivation and chase experiment for mitophagy: mitophagy flux response toward various stimulations

Mitophagy, a crucial mitochondrial quality control system for cellular stress adaptation, is a key focus in pathophysiology and drug discovery. Developing a simple and versatile mitophagy flux assay is vital for advancing our understanding of cellular responses. Addressing a gap in systematic method...

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Main Authors: Yoh Sugawara, Hiroyuki Morinaga, Jingyuan Chen, Yoshinori Kitagawa, Hiroki Ogata, Asiya Karim, Miu Kikuchi, Maryam Khan, Erica Yasuhara, Takahisa Goto, Joseph A. Jeevendra Martyn, Shingo Yasuhara
Format: Article
Language:English
Published: Taylor & Francis Group 2025-04-01
Series:BioTechniques
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Online Access:https://www.tandfonline.com/doi/10.1080/07366205.2025.2505357
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Summary:Mitophagy, a crucial mitochondrial quality control system for cellular stress adaptation, is a key focus in pathophysiology and drug discovery. Developing a simple and versatile mitophagy flux assay is vital for advancing our understanding of cellular responses. Addressing a gap in systematic methods, we employ the photoactivatable fluorescent protein mito-Kaede in C2C12 myocytes, demonstrating its remarkable versatility in quantifying mitophagy flux responses under various stimuli, including carbonyl cyanide m-chlorophenyl hydrazone (CCCP), TNF-α, lipopolysaccharide (LPS), and hypoxia. This study underscores the validity and distinctive advantages of the mito-Kaede assay through comparative analysis with conventional assays including Western blotting (WB), potentially providing valuable insights for both mitophagy flux analysis and drug development.
ISSN:0736-6205
1940-9818