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.
Faculty of Science, University of Split, Ruđera Boškovića 33, 21000, Split, Croatia.
Matilda
M
Šprung
School of Medicine, University of Split, Šoltanska 2, 21000, Split, Croatia.|Institute of Biochemistry II, Goethe University Medical School, Frankfurt, Germany.|Buchmann Institute for Molecular Life Sciences, 60590, Frankfurt, Germany.
Ivan
I
Dikic
School of Medicine, University of Split, Šoltanska 2, 21000, Split, Croatia. ivana.novak@mefst.hr.
Ivana
I
Novak
D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D063306:Mitophagy; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
C116838:BNIP3L protein, human; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D025521:Tumor Suppressor Proteins
Bnip3L/Nix; Flow cytometry; MitoTracker; Mitophagy; Mitophagy receptor