Quantifying Drp1-Mediated Mitochondrial Fission by Immunostaining in Fixed Cells

Methods Mol Biol. 2020:2159:197-204. doi: 10.1007/978-1-0716-0676-6_15.

Abstract

Dynamin-like protein 1 (Drp1) is the master regulator of mitochondrial fission. Drp1 translocates from the cytosol to the mitochondrial outer membrane to execute the scission process. Here we describe an immunofluorescence-based method to measure the mitochondrial translocation of Drp1 and quantify Drp1-related mitochondrial fission by labeling the mitochondrial import receptor subunit TOM20 in fixed cell culture.

Keywords: Confocal microscopy; Dynamin-related protein 1; Immunofluorescence staining; Mitochondrial fission; TOM20.

MeSH terms

  • Animals
  • Dynamins / genetics
  • Dynamins / metabolism*
  • Fibroblasts / metabolism
  • Fluorescent Antibody Technique* / methods
  • Gene Expression
  • Humans
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal / methods
  • Mitochondrial Dynamics*
  • Transfection

Substances

  • DNM1L protein, human
  • Dynamins