Extended Data Fig. 7. NLRP3 activators disrupt ETT but not overall TGN integrity.

a, Confocal images of Nlrp3 KO BMDMs treated with vehicle, 10 μM nigericin or 50 μM CL097 for 30 min. Cells were co-stained with antibodies against EEA1 and TGN38. DAPI was used to stain the nucleus. Magnifications of areas in dashed squares shown in the lower right corner. Scale bar: 10 μm. b, Pearson’s correlation coefficients of fluorescence of TGN38 and EEA1 in cells in experiments shown in panel a. Means ± SD, N = 3, n = 30 cells for control-treated, n = 35 cells for nigericin-treated and n = 35 cells for CL097-treated group. c, Confocal images of HeLa cells treated with vehicle, 10 μM nigericin for 40 min or 45 μg ml⁻¹ CL097 for 80 min. Cells were stained with antibodies against EEA1, TGN46 and GCC2. DAPI was used to stain the nucleus. Magnifications of areas in dashed squares shown in the lower right corner. Arrowheads indicate EEA1-positive endosomes containing TGN46. Scale bar: 10 μm. d, Pearson’s correlation coefficient of fluorescence of TGN46 and EEA1 (upper) and of TGN46 and GCC2 (lower) in experiments shown in panel c. Means ± SD, N = 3, n = 24 cells for control-treated, n = 18 cells for nigericin-treated and n = 21 cells for CL097-treated group. e, Confocal images of HeLa cells treated with vehicle, 10 μM nigericin for 40 min or 45 μg ml⁻¹ CL097 for 80 min. Cells were co-stained with antibodies against TGN46 and p230. DAPI was used to stain the nucleus. Magnifications of areas in dashed squares and shown in the lower right corner. Scale bar: 10 μm. f, Pearson’s correlation coefficient of fluorescence of TGN46 and p230 in experiments shown in panel e. Means ± SD, N = 3, n = 19 cells for control-treated group, n = 21 cells for nigericin-treated group and n = 22 cells for CL097-treated group. Data were analyzed with an unpaired two-sided t-test (b, d, f). Data shown in a, c, e are representative of at least three independent experiments.