Detection of Iron Depletion- and Hypoxia-Induced Mitophagy in Mammalian Cells

Mitochondrial autophagy or mitophagy is a process that selectively degrades mitochondria via autophagy. It is believed that mitophagy degrades damaged or unnecessary mitochondria and is important for maintaining mitochondrial homeostasis. To date, it is known that several stimuli can induce mitophagy. However, some of these stimuli (including iron depletion, hypoxia, and nitrogen starvation) induce mild mitophagy, which is difficult to detect by measuring the decrease in mitochondrial mass. Recently, we have successfully detected mitophagy induced under these conditions using mito-Keima as a reporter. In this chapter, we describe the protocols for induction and detection of iron depletion- and hypoxia-induced mitophagy using the mito-Keima-expressing cells.