Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Apr 19

PMID 37074639

Authors

William P Scott

Isabella Rauch

Affiliations

Soon will be listed here.

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.

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