Flow Cytometer Monitoring of Bnip3- and Bnip3L/Nix-Dependent Mitophagy

Mitochondria are organelles with numerous vital roles in cellular metabolism. Impaired or damaged mitochondria are degraded in autophagolysosomes in a process known as mitophagy. Given the fundamental role of mitophagy in maintenance of cellular homeostasis, methods and techniques with which to study this process are constantly evolving and emerging. So far, mitophagy flux was mostly monitored using fluorescently labeled LC3 protein on autophagosomal membrane and any of the labeled outer mitochondrial membrane proteins. However, this method is labor intensive, time consuming, and difficult to quantitatively validate due to the rapid mitochondrial turnover. Here, we describe a flow cytometry as a novel and promising quantitative method to monitor Bnip3- and Bnip3L/Nix-mediated mitophagy.