Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox

William P Scott; Isabella Rauch

doi:10.1007/978-1-0716-3040-2_3

Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox

Methods Mol Biol. 2023:2641:27-36. doi: 10.1007/978-1-0716-3040-2_3.

Authors

William P Scott¹, Isabella Rauch²

Affiliations

¹ Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, USA.
² Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, USA. rauchi@ohsu.edu.

PMID: 37074639
DOI: 10.1007/978-1-0716-3040-2_3

Abstract

Targeted activation of the NAIP-NLRC4 inflammasome has proven very useful in the study of pyroptosis. FlaTox and derivative LFn-NAIP-ligand cytosolic delivery systems offer a unique opportunity to interrogate both ligand recognition and downstream effects of the NAIP-NLRC4 inflammasome pathway. Here we describe how to stimulate the NAIP-NLRC4 inflammasome in vitro and in vivo. We describe experimental setup and specific considerations for treatment of macrophages in vitro and in vivo injections using a murine model of systemic inflammasome activation. The in vitro readouts of inflammasome activation propidium iodide uptake and lactate dehydrogenase (LDH) release as well as the in vivo readouts of hematocrit and body temperature measurement are described.

Keywords: FlaTox; Inflammasome; NAIP; NLRC4; Pyroptosis.

MeSH terms

Animals
Apoptosis Regulatory Proteins* / metabolism
Calcium-Binding Proteins / metabolism
Inflammasomes* / metabolism
Ligands
Macrophages / metabolism
Mice
Pyroptosis

Substances

Inflammasomes
Apoptosis Regulatory Proteins
Ligands
Calcium-Binding Proteins