Extended Data Fig. 2: NLRP3 activators trigger PI4P accumulation on endosomes in THP-1 cells and iBMDMs.

a, Confocal images of ASC KO THP-1 cells treated with vehicle (Ctrl), 15 µM nigericin or 50 µM CL097 for 30 min. After fixation, cells were co-stained with antibodies against PI4P and EEA1. DAPI was used to stain the nucleus. Magnifications of areas in dashed squares shown in the lower right corner. Scale bar: 10 μm. b, Quantification of PI4P levels on EEA1-positive structures in experiments shon in panel a. Means ± SD, N = 3, n = 16 cells for control-treated group; n = 19 for nigericin-treated group and n = 18 cells for CL097-treated group. c, Confocal images of ASC KO iBMDMs treated with vehicle or 15 μM nigericin for 30 min. Cells were co-stained with antibodies against PI4P and EEA1. Magnifications of areas in dashed squares shown in the lower left corner. Scale bar: 10 μm. d, Quantification of PI4P levels on EEA1-positive structures in experiments shown in panel c. Means ± SD, N = 3, n = 13 cells for control-treated group and n = 19 cells for nigericin-treated group. Data were analyzed with an unpaired two-sided t-test (b, d). Data shown in a, c are representative of at least three independent experiments.