Fig 4. MCC950/CRID3 inhibition of inflammasome responses in Nlrp3^A350V macrophages.

(A–C) BMDMs from tamoxifen-treated Nlrp3^A350V/+CreT− and Nlrp3^A350V/+CreT+ mice were left untreated (Ctrl) or treated with LPS in absence or presence of MCC950/CRID3. Supernatants were analyzed for IL-1β (A) and IL-18 (B) secretion, and lysates were immunoblotted for Casp1 and IL-1β (C). (D–F) BMDMs from tamoxifen-treated Nlrp3^A350V/+CreT− and Nlrp3^A350V/+CreT+ mice were left untreated (Ctrl) or primed with LPS and stimulated with ATP in absence or presence of MCC950/CRID3. Supernatants were analyzed for IL-1β (D) and IL-18 (E) secretion, and lysates were immunoblotted for Casp1 and IL-1β (F). (G–I) BMDMs from tamoxifen-treated Nlrp3^A350V/+CreT− and Nlrp3^A350V/+CreT+ mice were left untreated (Ctrl) or primed with LPS and stimulated with Nig in absence or presence of MCC950/CRID3. Supernatants were analyzed for IL-1β (G) and IL-18 (H) secretion, and lysates were immunoblotted for Casp1 and IL-1β (I). The numerical values underlying Fig 4A, 4B, 4D, 4E, 4G, and 4H can be found in S3 Data. Graphs show mean ± SD of triplicate wells and represent three independent experiments. BMDM, bone-marrow–derived macrophage; Casp1, caspase-1; Cre-ERT2, Cre recombinase-estrogen receptor fusion protein; CreT, Cre-ERT2 fusion gene; CRID, Cytokine Release Inhibitory Drug; Ctrl, control; IL, interleukin; LPS, lipopolysaccharides; LRR, leucine-rich repeat; NBD, nucleotide-binding domain; Nig, nigericin; NLR, NBD- and LRR-containing; NLRP3, NLR family, pyrin-domain–containing 3.