Extended Data Fig. 4: Neither retrograde trafficking nor endosomal maturation was required for PFO-mediated TGN remodeling or NLRP3 inflammasome activation.
a–c, Representative immunofluorescence images (a), quantification of TGN dispersion (b) and NLRP3 recruitment (c) in HeLa cells stably expressing NLRP3-GFP pre-treated with DMSO (solvent control), 25 µM retro-2cycl or 20 µM EGA for 1 h before addition of 0.90 nM rPFO or not (Mock) for 80 min. Scale bar, 10 μm. Areas containing TGN structures were measured with ImageJ (n = 40 cells per condition; mean ± s.d.; two-sided t-test; NS, not significant). NLRP3 recruitment was quantified from 100 cells (n = 3; N.D., not detectable). Representative from 3 independent experiments. d, Immunoblots of HEK293T cells stably expressing NLRP3, ASC, and caspase-1 (Casp1) pre-treated with DMSO (solvent control), 25 µM retro-2cycl, or 20 µM EGA for 1 h before addition of 0.90 nM rPFO 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). Representative from 2 independent experiments.
