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  • Immunofluorescence microscopy-based assessment of cytosolic DNA accumulation in mammalian cells.

Immunofluorescence microscopy-based assessment of cytosolic DNA accumulation in mammalian cells.

STAR protocols (2021-05-28)
Ai Sato, Aitziber Buque, Takahiro Yamazaki, Norma Bloy, Giulia Petroni, Lorenzo Galluzzi
摘要

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).

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TS/A Mouse Mammary Adenocarcinoma Cell Line, TS/A mouse mammary adenocarcinoma cell line is a well characterized and highly published model for breast cancer research.