Extended Data Fig. 1 |. Comparative analysis of IL-1 cytokine cleavage across mammals.

a, 10 μg of indicated proteins were separated by SDS-PAGE and stained with InstantBlue. b-k, In vitro cleavage of pro-forms of indicated human IL-1 family cytokines by caspase-4, caspase-1 and caspase-5. Immunoblots are representative of three independent repeats. Asterisk marks signal due to cross-reaction of primary antibody with His-tagged p20 subunit of the caspase. l,m, Immunoblots showing in vitro cleavage of human pro-IL-1β by human caspase-1 and caspase-5. Asterisk marks signal due to cross-reaction of primary antibody with His-tagged p20 subunit of the caspase. n-q, Immunoblots showing in vitro cleavage of murine pro-IL-18 by human caspase-1 and caspase-4, and murine caspase-1 and caspase-11. r–v, Immunoblots showing in vitro cleavage of pro-IL-18 from indicated mammalian species by caspase-4 homolog from the same species. w, x, Immunoblot and quantification of in vitro cleavage of human pro-IL-18 mutants by murine caspase-11. n=4 biological replicates for caspase-4 and n=3 biological replicates for caspase-11. All immunoblots are representative of at least three biological replicates. Bars and error bars represent mean ± SEM. Statistical significance was determined by unpaired, two-sided student’s t-test. For gel source data, see Supplementary Figure 1.