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Methods Mol Biol. 2017;1656:131-142. doi: 10.1007/978-1-4939-7237-1_7.

Methods to Visualize MAVS Subcellular Localization.

Author information

1
Department of Molecular Genetics and Microbiology, Duke University Medical Center, 213 Research Dr., Box 3053, Durham, NC, 27710, USA.
2
Department of Molecular Genetics and Microbiology, Duke University Medical Center, 213 Research Dr., Box 3053, Durham, NC, 27710, USA. stacy.horner@duke.edu.
3
Department of Medicine, Duke University Medical Center, 213 Research Dr., Box 3053, Durham, NC, 27710, USA. stacy.horner@duke.edu.

Abstract

The mitochondrial antiviral signaling (MAVS) protein is a central adaptor protein required for antiviral innate immune signaling. To facilitate its roles in innate immunity, MAVS localizes to multiple intracellular membranous compartments, including the mitochondria, the mitochondrial-associated ER membrane (MAM), and peroxisomes. Studies of MAVS function therefore often require an analysis of MAVS localization. To detect MAVS protein on intracellular membranes, biochemical fractionation to isolate MAMs, mitochondria, or peroxisomes can be used. Further, immunofluorescence with antibodies against specific membrane markers can be used to visualize MAVS distribution throughout the cell. Here, we describe the biochemical fractionation and immunofluorescence protocols used to detect MAVS subcellular localization.

KEYWORDS:

Endoplasmic reticulum; Fractionation; Immunofluorescence; Interferon; MAM; MAVS; Mitochondria; Peroxisomes

PMID:
28808966
PMCID:
PMC6103534
DOI:
10.1007/978-1-4939-7237-1_7
[Indexed for MEDLINE]
Free PMC Article

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