Extended Data Fig. 6: Sulfhydryl-containing lipid chains are linked by disulfide bonds.
From: Cytosolic CTH senses bacterial lipoproteins and drives noncanonical inflammasome activation
a, Immunoblots of caspase-11 cleavage after incubation with lipid A (50 ng/μl), SDG (10 ng/μl) or DDG (10 ng/μl). b, Immunoblots of BMDMs from Cth+/+ and Cth−/− mice primed with 1 μg/ml Pam3CSK4 overnight before transfected with DOTAP alone or together with 10 μg/ml SDG or DDG for 16 h. c, Surface plasmon resonance analysis of caspase-11 binding to lipid A, SDG or DDG, GST as control. d, Immunoblots of BMDMs from wild-type, Casp11−/− and Casp1−/− mice primed with 1 μg/ml Pam3CSK4 overnight before infected with live S. aureus at an MOI of 10 for 6 h. Cells cultured in fresh medium containing gentamycin (100 μg/ml) with or without 0.1 mM H2O2 for 16 h. e, Quantification of pyroptosis in BMDMs from wild-type, Casp11−/− and Casp1−/− mice treated as in d. ELISA assays of IL-1β in supernatants, LDH release assays of cell death in supernatants and ATP assays of cell viability in cell pellets. f, Quantification of pyroptosis in BMDMs from wild-type, Casp11−/− and Casp1−/− mice primed with 1 μg/ml Pam3CSK4 overnight before infected with live S. aureus at an MOI of 10 for 6 h. Cells cultured in fresh medium containing gentamycin (100 μg/ml) with or without PAG (0.6 mM) for 16 h. LDH release assays of cell death in supernatants and ATP assays of cell viability in cell pellets. In e and f, data are pooled from three independent experiments (n = 3) and shown as means ± SD. Unpaired two-tailed Student’s t-test was used. Exact P values are indicated in the figures. Data are representative of three independent experiments with similar results in a-f.
