Extended Data Fig. 3: Endogenous NLRP3 is recruited to dTGN in primary macrophages.

a, Substantial TGN disassembly occurred at early time points in wild-type BMDMs. Cells were primed with LPS (50 ng ml⁻¹) for 3 h, followed by stimulation with nigericin (10 μM) or ATP (5 mM) for the indicated time, and immunostained for TGN38. Right, to quantify TGN disassembly, the numbers of TGN structures not connected with each other for each cell were quantified from 100 randomly selected cells and grouped as shown. b, c, Nigericin-induced cleavage of caspase-1 and IL-1β did not occur until 15 min (for nigericin) or 10 min (for ATP) after stimulation in wild-type BMDMs. Cells were treated as in a before lysates were collected for immunoblotting. d, Endogenous NLRP3 aggregation on dTGN could be detected as early as 10 min after nigericin treatment in ASC-deficient BMDMs. Cells were primed with LPS (50 ng ml⁻¹) for 3 h, followed by nigericin (10 μM) treatment for 0, 10 or 30 min before imaging. Cells with NLRP3 puncta on dTGN were quantified from 100 cells (n = 3, mean ± s.d., two-sided t-test).