Extended Data Fig. 10: Model: NLRP3 aggregation on dTGN through PtdIns4P binding is a common cellular signal essential for the inflammasome activation by diverse stimuli.

Centre, under basal conditions, NLRP3 is diffused in the cytosol and TGN (red) remains as a single cluster attached to medial- and cis-Golgi (purple). Left, when cells are stimulated with K⁺ efflux-dependent stimuli such as nigericin or ATP, TGN is disassembled into multiple dispersed structures (dTGN), whereas cis- and medial-Golgi stacks remain intact. These stimuli also trigger K⁺ efflux, which helps recruit NLRP3 to dTGN via ionic bonding between negatively charged PtdIns4P on dTGN membranes and the positively charged polybasic region of NLRP3. Right, when cells are stimulated with K⁺ efflux-independent stimuli such as imiquimod and CL097, TGN is also disassembled, although more drastically. PtdIns4P is partially separated from other TGN compartments; some may be enriched on the plasma membrane. NLRP3 is then recruited to these PtdIns4P-containing microdomains through its polybasic region. For both types of stimulation, dTGN serves as a scaffold for NLRP3 to aggregate in the form of multiple puncta, which then interact with ASC to activate the downstream signalling cascade. This model shows that aggregation of NLRP3 on dTGN through PtdIns4P binding is a common cellular signal that is essential for its activation by diverse stimuli.