Immunofluorescence microscopy-based assessment of cytosolic DNA accumulation in mammalian cells
Ai Sato, Aitziber Buqué, Takahiro Yamazaki, Norma Bloy, Giulia Petroni, Lorenzo Galluzzi
Abstract
Here, we describe an immunofluorescence (IF) microscopy-based approach to quantify cytosolic double-stranded DNA molecules in cultured eukaryotic cells upon the selective and specific permeabilization of plasma membranes. This technique is compatible with widefield microscopy coupled with automated image analysis for mid- to high-throughput applications and high-resolution confocal microscopy for subcellular assessments and co-localization studies. In addition to enabling single-cell and subcellular resolution, this approach circumvents most constraints associated with alternative approaches based on subcellular fractionation. For complete use and execution of this protocol, please refer to Yamazaki et al. (2020).