Extended Data Fig. 4: ADCY7 enzymatic activity is required for inflammasome activation.
From: Eukaryotic ADCY7 catalyzes the production of c-di-AMP to activate the NLRP3 inflammasome
a–c, Adcy7–/– BMDM cells were rescued with wild-type or enzymatic-mutant ADCY7 without transmembrane domains. Cells were stimulated with 1 μM control ODN or CpG-ODN for 6 h. (a) Cell culture supernatants were collected for an ELISA assay to determine the secreted IL-1β protein levels. (b) Cytotoxicity was determined by LDH release assay in cell culture supernatants. (c) Cell viability was determined by an ATP quantification assay in cell pellets. d–g, Macrophages derived from human peripheral blood mononuclear cells were infected with lentiviruses carrying Cas9 and an sgRNA against human ADCY7. (d) The efficiencies of gene deletion were examined through immunoblotting. ADCY7+/+ and ADCY7–/– human macrophages were stimulated with 1 µg/ml E. coli genomic DNA for 6 h. (e) Cell culture supernatants were collected for an ELISA assay to determine the secreted IL-1β protein levels. (f) Cytotoxicity was determined by LDH release assay in cell culture supernatants. (g) Cell viability was determined by an ATP quantification assay in cell pellets. In a–c and e–g, data were shown as means ± SD (n = 4, biological replicates). Two-tailed Student’s t-test were used.
