Extended Data Fig. 3: In vitro activation and modulation of the Nlrp10 inflammasome.
From: Epithelial Nlrp10 inflammasome mediates protection against intestinal autoinflammation
(a) IL-18 ELISA of supernatants of human ASCTagBFP HEK cells (HEKASC) and human ASCTagBFP HEK cells expressing human Nlrp10mCitrine (HEKASC_Nlrp10) transfected with human myc- caspase-1, and pro-IL-18, and treated with m-3M3FBS and a panel of toll-like receptor (TLR) agonists, including the TLR2 agonist HKLM, TLR2/6 agonist FSL1, TLR4 agonist LPSEK, TLR5 agonist FLAST, and TLR3 agonist poly(I:C) (LMW: low molecular weight, HMW: high molecular weight) (n = 3) (Unpaired t-test, P values are *P = 0.018948, **P = 0.005017, ***P = 0.000883). (b) Human ASCTagBFP HEK cells (HEKASC) and human ASCTagBFP HEK cells expressing human Nlrp10mCitrine (HEKASC_Nlrp10) and HEK cells expressing human Nlrp10mCitrine (HEKNlrp10) immunoprecipitated using anti-GFP beads to pull down Nlrp10 and immunoblotted with anti-GFP and anti-ASC. (c) IL-18 (Unpaired t-test, ****P < 0.0001) and (d) IL-1β (Unpaired t-test, ***P = 0.0001, **P = 0.005, ****P < 0.0001) ELISA of supernatants of Aim2−/−CRISPR immortalized mouse macrophages stably expressing full-length human Nlrp10- mCitrine (MacsAim2−/−_hNlrp10), mouse Nlrp10-mCitrine (MacsAim2−/−_mNlrp10), human Nlrp10-PYD (only)-mCitrine (MacsAim2−/−_PYD), or human Nlrp10-NACHT (only)-mCitrine (MacsAim2−/−_NACHT), treated with HMW poly(I:C), m-3M3FBS either alone or in combination (n = 3). (e) IL-1β ELISA of supernatants of differentiated organoids derived from the distal colons of Nlrp10loxP/loxPCMV-Cre+ and Nlrp10loxP/loxP littermates treated with vehicle, HMW poly(I:C), or both HMW Poly(I:C) and m-3M3FBS (n = 6, results pooled from 2 independent experiments; n = 3 in the positive control group). CMV: Cytomegalovirus. Recombinant IL-1β was used as the positive control. Mean ± SEM. *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001. Statistical test used in (a), (c-d) was two-sided.
