Subcellular location of NLRP3 is not essential, but clustering on different scaffolds is. This explains how NLRP3 can react to so many diverse stress signals.
9/9

Surprisingly, this activation could not be blocked by MCC950, suggesting that clustering of NLRP3 induces rearrangements of inactive NLRP3 leading to active NACHT-based oligomer formation.
8/9

To test if lipid membranes are truly dispensable, we fused NLRP3 to TDP43, a protein that forms cytosolic condensates. That alone was enough to trigger inflammasome assembly without any external stimulus.
7/9

We showed that NLRP3 can be activated from different types of scaffolds not just membranes. Variants targeted to the Golgi (GOLGB1) or centrosome (PACT) triggered IL-1β release after priming alone. Both formed multiple spots in the cell, resembling activated NLRP3.
6/9

The polybasic segment in NLRP3 is highly conserved and known to mediate lipid binding. We mutated this region (4xA) to test if it is essential for activation. Interestingly, membrane-enriched variants still triggered IL-1β secretion, whereas cytosolic 4xA failed.
5/9

All designed variants formed a functional inflammasome judged by ASC speck formation and their activation was K+ efflux-dependent.
4/9

We engineered NLRP3 to localize to different organelles: ER, Golgi, plasma membrane, lysosomes, mitochondria and peroxisomes. Surprisingly, all location-restricted variants supported IL-1β release when stimulated with different activators.
3/9