Monitoring Inflammasome Activity Through ASC Speck Formation in THP-1 ASC-GFP Cells

Inflammasome activation and signaling are hallmarks of immune activation and cellular response. Understanding the complex mechanisms involved in the formation of the NLRP3 inflammasome is vital to accurately evaluating potentially anti-inflammatory molecules. The oligomerization of ASC is a well-documented indicator of inflammasome activation, and ASC-GFP allows the visualization of these "specks." Here, we detail a higher throughput methodology for monitoring inflammasome activation and screening potential anti-inflammatory compounds utilizing an Incucyte® live cell imager. It provides a platform to observe cell activation and cell death in real-time. Cells can be visualized from initial plating through to inflammasome-induced cell death, granting insights into inflammasome activation kinetics and previously unknown toxicities.