Fig. 7. NLRP11 is necessary for IL-1β release in CAPS.

a, qPCR of NLRP3 and NLRP11 mRNA levels from THP-1 cells primed with LPS (200 ng ml⁻¹) as indicated, presented as fold expression to average mRNA levels present in uninduced cells (n = 3, mean ± s.d.); *P < 0.0001, **P = 0.0103, ***P = 0.0145, ****P = 0.0037. b, IL-1β ELISA of SN from Cas9^Ctrl, NLRP11^KO, NLRP3^KO, ASC^KD, NLRP11^Flag(lo) and NLRP11^Flag(hi) cells primed with LPS (200 ng ml⁻¹, 4 h) and primed and activated with nigericin (5 μM, 30 min) (n = 3, mean ± s.d.); *P = 0.0006; **P = 0.0012, ***P < 0.0001, ****P = 0.0021. c, Fluorescence microscopy of EGFP and DAPI in HEK293^NLRP3-EGFP and HEK293^{NLRP3R260W-EGFP} cells transiently cotransfected with empty vector (Ctrl) or NLRP11; scale bars, 100 μm (left), and quantification of NLRP3 oligomer⁺ cells per view (right) (Ctrl: n = 3, NLRP11: n = 5, mean ± s.d.); *P < 0.0001; **P = 0.029. d, Confocal microscopy of DAPI and NLRP3 immunostained PMA-differentiated Cas9^Ctrl, NLRP11^KO, NLRP11^Flag(lo) and NLRP11^Flag(lo) and NLRP3^KO cells stably expressing NLRP3^R260W left untreated (Ctrl) or primed with LPS (200 ng ml⁻¹, 4 h); scale bars, 50 μm (left), and quantification of NLRP3 oligomer⁺ cells per view (right) (n = 5, mean ± s.d.); *P = 0.0017; **P = 0.0046, ***P < 0.0001, ****P = 0.0153, *****P = 0.0003, ******P = 0.0001, *******P = 0.0427, ********P = 0.0002. e, IL-1β ELISA of SNs from NLRP11^KO and NLRP11^Flag cells stably expressing NLRP3^R260W left untreated (Ctrl) or primed with Pam3CSK4 (1 μg ml⁻¹, 2 h) (n = 3, mean ± s.d.). *P < 0.0001.

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