Fig. 5. NLRP3 activators disrupt ETT.

a, Internalization of StxB in HeLa cells. Cells were treated or not treated with 10 μM nigericin. Cy3-conjugated StxB was added into culture medium 5 min after the addition of nigericin. Cells were fixed at 40 min after incubation with Cy3-conjugated StxB. Cells were stained with antibodies against GCC1 and EEA1. DAPI was used to stain the nucleus. Magnifications of areas in dashed squares are shown in the lower right corner. Arrowheads indicate StxB retained on EEA1-positive endosomes. Scale bar, 10 μm. b, Internalization of CI-MPR antibody in HeLa cells. Cells were treated or not treated with 10 μM nigericin or 45 μg ml⁻¹ CL097. An antibody recognizing CI-MPR was added into the culture medium 5 min after nigericin or 20 min after CL097 treatment. Cells were fixed 40 min after the incubation with the CI-MPR antibody and were stained with an antibody against EEA1. DAPI was used to stain the nucleus. Magnifications of areas in numbered dashed squares are shown in separate numbered images. Scale bar, 10 μm. c, Quantification of StxB levels on endosomes and Golgi in experiments as described for a. Mean ± s.d., N = 3, n = 43 cells for control (ctrl) group and n = 40 cells for nigericin-treated group. d, Quantification of CI-MPR levels on endosomes in experiments in b. Mean ± s.d., N = 3, n = 47 cells for control group; n = 48 cells for nigericin-treated group and n = 44 for CL097-treated group. Data were analyzed with an unpaired two-tailed t-test (c, d). Data shown in a and b are representative of at least three independent experiments.