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Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox.

William P Scott1, Isabella Rauch2

  • 1Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, USA.

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Summary
This summary is machine-generated.

This study details methods for activating the NAIP-NLRC4 inflammasome using FlaTox systems. It outlines in vitro and in vivo experimental procedures and key measurements for studying pyroptosis.

Keywords:
FlaToxInflammasomeNAIPNLRC4Pyroptosis

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Area of Science:

  • Immunology
  • Cellular Biology

Background:

  • The NAIP-NLRC4 inflammasome is crucial for studying pyroptosis.
  • Targeted activation is key to understanding its role.

Purpose of the Study:

  • To describe methods for stimulating the NAIP-NLRC4 inflammasome in vitro and in vivo.
  • To provide guidance on experimental setup and considerations for macrophage treatment and systemic inflammasome activation models.

Main Methods:

  • Utilized FlaTox and LFn-NAIP-ligand delivery systems for cytosolic delivery.
  • Described in vitro stimulation of macrophages and in vivo injections in a murine model.
  • Detailed measurement of propidium iodide uptake, LDH release, hematocrit, and body temperature.

Main Results:

  • Established protocols for targeted NAIP-NLRC4 inflammasome activation.
  • Demonstrated feasibility of in vitro and in vivo stimulation and readout measurements.
  • Provided a framework for interrogating inflammasome ligand recognition and downstream effects.

Conclusions:

  • The described methods enable robust investigation of the NAIP-NLRC4 inflammasome pathway.
  • These techniques facilitate the study of pyroptosis and related inflammatory responses.
  • This work offers a valuable resource for researchers in immunology and cell biology.