Extended Data Fig. 6: GSDMA-NT produced by SpeB cleavage is sufficient to initiate pyroptosis, but does not injure bystander cells.
From: Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis
a, Coomassie Blue-stained SDS-PAGE gel showing purified recombinant GSDMA, GSDMA-NT and GSDMA-CT. b–d, Equal amounts of recombinant full-length GSDMA, GSDMA-NT (1–246aa), GSDMA-CT, full-length GSDMD, Caspase-11 or full-length GSDMD plus Caspase-11 were electroporated into 293T cells respectively, followed by cell morphology observation by phase-contrast microscopy (b), cell viability analysis by CellTiter-Glo luminescent assay (c), and cell death assessment by PI uptake (d). e–g, Equal amounts of recombinant of full-length GSDMA, GSDMA-NT (1–246aa), GSDMA-CT, full-length GSDMD, Caspase-11 or full-length GSDMD plus Caspase-11 were respectively added directly into cell culture medium, followed by cell morphology observation by phase-contrast microscopy (e), cell viability analysis by CellTiter-Glo luminescent assay (f), and cell death assessment by PI uptake (g). Pyroptotic cells form large ballooning bubbles; Scale bar: 20 μm. Graphs show mean ± s.d. of triplicate wells (c, f) or mean ± s.e.m. of quadruplicate wells (d, g). c, Two-tailed Student’s t-test. Data are representative of at least three independent experiments. For gel source data, see Supplementary Fig. 1.
