Extended Data Fig. 2: Multiple CDCs induced TGN remodeling and NLRP3 recruitment.
a, Coomassie blue staining for CDCs purified from E. coli. Arrows indicate the CDC proteins. Representative from at least 3 independent experiments. b, Representative LDH release assay of HeLa cells incubated with the indicated CDCs for 80 min (n = 3 wells per condition; mean ± s.d.). Representative from 2 independent experiments. c, Immunoblots of HEK293T cells stably expressing NLRP3, ASC, and caspase-1 (Casp1) incubated with 10 μM nigericin, 0.18/0.54/1.8/5.4 nM rALO, or 1.8/5.4/10.8/14.4 nM rSLO for 40 min. pro-Casp1 has two bands in this cell line because it was expressed as zeocinr-F2A-Casp1 (upper band) before ribosomal skipping to release pro-Casp1 (lower band). SE, short exposure; LE, long exposure. Representative from 3 independent experiments. d, Representative immunofluorescence images (left) and quantification of TGN dispersion and NLRP3 recruitment (right) in HeLa cells stably expressing NLRP3-GFP incubated with 0.18/0.54//0.90/1.8 nM rALO or not (Mock) for 80 min. Representative images for mock treatment and 1.8 nM rALO treatment are shown. Scale bar, 10 μm. Areas containing TGN structures were measured with ImageJ (n = 40 cells per condition; two-sided t-test). NLRP3 recruitment was quantified from 100 cells (n = 3; N.D., not detectable). Representative from at least 3 independent experiments. e, As in d, except cells were incubated with 5.4/10.8/14.4/18 nM rSLO or not (Mock). Representative images for mock treatment and 14.4 nM rSLO treatment are shown. Scale bar, 10 μm. Representative from at least 3 independent experiments.
