Fig. 2.

Splenic DCs express NLRP3 inflammasome components. Splenic DCs were isolated from mice at 24 h after CLP or sham operation. A, Representative flow cytometry plots showing the gating strategy used to distinguish DCs and macrophages using the markers of CD11c, CD11b, MHC-II, Flt3 (CD135), CD115. CASP-1 (FAM-FLICA) staining of primary splenic CD11c⁺CD11b^intMHC-II^hiCD135⁺CD115⁻ DCs was analyzed by flow cytometry. The numbers of CD11c⁺CD11b^intMHC-II^hiCD135⁺CD115⁻ DCs, the parents of CD11c⁺ MHC-II^hiCD135⁺ DCs and the parents of CASP-1⁺ DCs are shown on the right. B, The DC pyroptosis rate was assessed by flow cytometry, and DCs were labeled with active CASP-1 (FAM-FLICA) and 7-AAD. Representative flow cytometry plots are shown on the left, and quantitative analysis of FLICA⁺ 7AAD⁺ DCs is shown on the right (n = 6). C, The protein expression levels of CASP-1, GSDMD, and IL-1β were determined by Western blotting. The values are protein levels relative to the β-actin level. The data shown are representative of 3 independent experiments. Statistical significance: ^*P < 0.05 versus the sham or control group