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Protocol for induction and characterization of microglia extracellular traps in murine and human microglia cells.

STAR protocols (2021-08-07)
Ishan Agrawal, Nidhi Sharma, Shivanjali Saxena, S Arvind, Debayani Chakraborty, Debarati Bhunia Chakraborty, Deepak Jha, Surajit Ghatak, Sridhar Epari, Tejpal Gupta, Sushmita Jha
RESUMEN

Extracellular traps (ETs) are composed of decondensed chromatin and are embedded with various antimicrobial proteins like myeloperoxidase and histones. Recently, we reported that dopamine (DA) induces ETs in BV2 microglia cell line and primary adult human microglia in a manner independent of cell death, reactive oxygen species, and actin polymerization. This protocol details how to characterize DA-induced ETs in BV2 microglia and human microglia. The protocols for characterization of ETs may also be used for other adherent cell lines. For complete details on the use and execution of this protocol, please refer to Agrawal et al. (2021).

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Sigma-Aldrich
Dopamina hydrochloride
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N-Acetyl-L-cysteine, BioReagent, suitable for cell culture
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Cytochalasin D, Ready Made Solution, from Zygosporium mansonii, 5 mg/mL in DMSO
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Anti-DNA/Histone H1 Antibody, Chemicon®, from mouse
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Anti-Neutrophil Myeloperoxidase antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution