Extended Data Fig. 1: Biochemical characterization of IpaH7.8–GSDMB interaction.
From: Structural mechanisms for regulation of GSDMB pore-forming activity
a, Assay of GSDMB (or GSDMD) degradation by IpaH7.8. Flag-IpaH7.8 (WT or C357S) was co-expressed with GSDMB or GSDMD in 293T cells. Cell lysates were subjected to immunoblotting analyses. b, c, Co-immunoprecipitation assay of IpaH7.8 with human gasdermins (GSDMs). IpaH7.8C357S mutant (b) or IpaH7.8LRR (c) was co-expressed with an indicated Flag-EGFP-tagged GSDM in 293T cells. Cell lysates were subjected to anti-Flag immunoprecipitation followed by immunoblotting. d, f, Gel-filtration chromatography analyses of complex formation between IpaH7.8 or IpaH7.8LRR and GSDMB or GSDMD (FL or the C-terminal domain). Elution profiles along with SDS-PAGE analyses of complex elution fractions are shown. e, SPR profiles of IpaH7.8 binding to GSDMB or human or mouse GSDMD. g, h, Gel-filtration chromatography analyses of complex formation between IpaH7.8 (WT or indicated GSDMB-binding site mutants) and GSDMB (WT or indicated IpaH7.8-binding site mutants). The binding-site mutations in IpaH7.8 and GSDMB were identified from the complex crystal structure. All data are representative of three independent experiments. See Supplementary Fig. 1 for gel source data.
