Tuft cell inflammasome expression and activation effects. (A) Tuft cell numbers are not affected by genotype or tamoxifen diet. Colony C57BL/6 or Nlrc4^–/–Pou2f3-cre^ERT2+ and iNLRC4 Pou2f3-cre^ERT2+ littermates were fed normal chow or tamoxifen chow for 7 days and the percentage tuft cells (CD24⁺SiglecF⁺) among EpCam⁺ cells was determined by flow cytometry (n = 3–7). (B) Percentage GFP positive (iNLRC4 expressing) tuft- and non-tuft cells from mice treated as in A (n = 3–4). (C) Epithelial monolayers were treated with FlaTox alone or FlaTox and BABTA-AM or Disulfiram and supernatants were analyzed for eicosanoid release. n = 4–5 combined data from two experiments; ****, P < 0.0001. (D and E) iNLRC4-Pou2f3-Cre^ERT2 and Nlrc4^−/−Pou2f3-Cre^ERT2 littermate controls were retro-orbitally injected with two FlaTox dosages (LFn-FlaA [0.1 µg/g] and PA [0.2 µg/g]) 48 h apart and tissues were harvested 24 h after the last injection. IL-18 (D) and IL-1α (E) protein levels within the small were quantified by ELISA, data pooled from multiple experiments (n = 12–18).