Extended Data Fig. 1. Galectin-1 is released as a consequence of inflammasome activation.

a, Experimental design for profiling caspase-11-mediated release of DAMPs by PF2D-based proteomics. WT and Casp11^−/− BMDMs were infected with EHEC (MOI=50) for 16 h and supernatants were harvested and concentrated and processed on the Beckman Coulter ProteomeLab PF2D platform. b, Galectin-1 secretion by Pam3CSK4-primed WT and Lgals1^−/− BMDMs stimulated with EHEC (MOI=50) or poly(dA:dT) transfection for 16 h or 10 μM nigericin for 1 h. c, Immunoblot of galectin-1 (14.5 kDa) in the supernatant and lysates of BMDMs stimulated as in (b). d, IL-1α secretion by Pam3CSK4-primed WT, Casp11^−/−, and Casp1^−/− BMDMs stimulated with 10 μM nigericin for 1 h. e, Immunoblot of caspase-11 and β-actin in lysates of MS1 endothelial cells transduced with LentiCRISPR V2 plasmid expressing control (GFP) sgRNA or caspase-11 sgRNA. f, Immunoblot of caspase-4 and β-actin in lysates of HeLa cells transduced with LentiCRISPR V2 plasmid expressing control (GFP) sgRNA or caspase-4 sgRNA. g, Immunoblot of GSDMD in the lysates of Pam3CSK4-primed WT BMDMs infected with EHEC or S. flexneri (MOI=50) for 16 h or 10 μM nigericin for 1 h in the presence or absence of 50 mM glycine. h, Galectin-1 release from liposomes packaged with galectin-1 and incubated with either 1 μg recombinant active caspase-11, 2 μg recombinant gasdermin D, both caspase-11 and gasdermin D, or 0.1% Triton X-100. Data are presented as mean ± SEM of one experiment representative of two (b,h; h is the replicate for the Fig. 2i). Combined data from two independent experiments (d) are shown as mean ± SEM. Immunoblots (c,e–g) are representative of two independent experiments.