Extended Data Fig. 10: SpeB cleaves mouse Gsdma1 and releases its N-terminal pyroptosis-inducing activity.
From: Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis
a, Whole cell lysates of 293T cells transfected with the indicated plasmids were collected and subjected to immunoblot analysis with the indicated antibodies. b, Coomassie Blue-stained SDS-PAGE gel showing in vitro cleavage of recombinant WT Gsdma1 or mutant Gsdma1 I246N/Q247E (1 μM) incubated with or without recombinant WT SpeB (100 nM) for 0.5 h. c, Whole cell lysates of 293T cells transfected with the indicated plasmids were collected and subjected to immunoblot analysis. d, Leakage of PC-PE liposomes containing additional PS or CL was monitored in real-time by terbium (Tb3+) fluorescence after incubation with recombinant gasdermins proteins in the presence or absence of recombinant SpeB as indicated. e, f, 293T cells transfected with the indicated plasmids were analysed by phase-contrast microscopy (e), LDH release (f). Arrowheads indicate pyroptotic cells; Scale bar: 10 μm. Graphs show mean ± s.d. of triplicate wells. f, One-way ANOVA. Data are representative of at least three independent experiments. For gel source data, see Supplementary Fig. 1.
