Fig. 1: Loss NLRP6 augments liver disease progression in NEMO^∆hepa mice.

a Macroscopic appearance of NEMO^∆hepa and NEMO^∆hepa/Nlrp6^−/− livers at 52 weeks of age. b Quantification of liver tumors with a diameter larger than 1 cm in NEMO^∆hepa (n = 13) and NEMO^∆hepa/Nlrp6^−/− (n = 8) livers; unpaired two-tailed Student’s t test, 95% CI −2.239 to −0.107, p = 0.0328. c Serum ALT and GLDH levels of 52-week-old NEMO^∆hepa (n = 12), NEMO^∆hepa/Nlrp6^−/− (n = 8)) and respective controls (WT (n = 6), Nlrp6^−/− (n = 6)); one-way ANOVA with Tukey’s multiple comparisons test (ALT: WT vs. NEMO^∆hepa, 95% CI −708.8 to −141.2, p = 0.0018; WT vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −1023 to −410.1, p = <0.0001; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −519.1 to −0.93, p = 0.0489, GLDH: WT vs. NEMO^∆hepa, 95% CI −339.4 to −127.3, p < 0.0001; WT vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −494.2 to −265.1, p = <0.0001; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −243.1 to −49.49, p = 0.049). d Representative pictures of H&E-stained liver sections, immunohistochemical (IHC) staining of CD45 and Sirius red stained liver sections of 52-week-old WT, Nlrp6^−/−, NEMO^∆hepa, and NEMO^∆hepa/Nlrp6^−/− livers, representative of 2 independent experiments; Scale bar: 100 µm. e RT-qPCR analysis of pro-fibrotic mRNA expression (TGFβ, Col1a1) in whole liver of NEMO^∆hepa (n = 12), NEMO^∆hepa/Nlrp6^−/− (n = 11) and respective controls (WT (n = 6), Nlrp6^−/− (n = 8)); one-way ANOVA with Sidak’s multiple comparisons test (TGFb: WT vs. NEMO^∆hepa, 95% CI −0.661 to 0.8128, p = ns; Nlrp6^−/− vs. NEMO^∆hepa/Nlrp6^−/−,95% CI –1.418 to −0.0483, p = 0.0328; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −1.252 to −0.0219, p = 0.0405; Col1a1: WT vs. NEMO^∆hepa, 95% CI −13.08 to 10.38, p = ns; Nlrp6^−/− vs. NEMO^∆hepa/Nlrp6^−/−,95% CI –26.78 to −3.865, p = 0.006; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −24.53 to −3.950, p = 0.004). f Representative pictures of immunofluorescence (IF) stainings of CD11b and CD8 of NEMO^∆hepa, NEMO^∆hepa/Nlrp6^−/− and respective controls (WT, Nlrp6^−/−). Nuclei were counterstained with DAPI, representative of 2 experiments; Scale bar: 100 µm. g RT-qPCR analysis of inflammatory mRNA expression (TNFα, IL-1β,Tlr4) in whole liver of NEMO^∆hepa (n = 12), NEMO^∆hepa/Nlrp6^−/− (n = 11) and respective controls (WT (n = 4), Nlrp6^−/− (n = 5)); one-way ANOVA with Sidak’s multiple comparisons test (Tnfa: WT vs. NEMO^∆hepa, 95% CI −2.37 to 0.6196, p = ns; Nlrp6^−/− vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI –4.04 to −0.806, p = 0.0015; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −2.505 to −0.006, p = 0.023; IL-1β: WT vs. NEMO^∆hepa, 95% CI −1.45 to 1.72, p = ns; Nlrp6^−/− vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI –2.90 to −0.1197, p = ns; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −2.601 to −0.1138, p = 0.047; Tlr4: WT vs. NEMO^∆hepa, 95% CI −5.62 to 4.156, p = ns; Nlrp6^−/− vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI –6.61 to −2.71, p = ns; NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −8.989 to −1.323, p = 0.005). h Immunoblot analysis of liver protein extracts from 52-week-old mice of all indicated genotypes for JNK, p-JNK and GAPDH as loading control; representative of 2 experiment. i Histological quantification of Cleaved Caspase 3 positive cells per viewfield; NEMO^∆hepa (n = 17) and NEMO^∆hepa/Nlrp6^−/− (n = 24) viewfields one-way ANOVA with Tukey’s multiple comparisons test (NEMO^∆hepa vs. NEMO^∆hepa/Nlrp6^−/−, 95% CI −8.141 to −3.410, p < 0.0001). j Representative pictures of IF staining of KI67. Nuclei were counterstained with DAPI, representative of 2 experiments; Scale bar: 100 µm. All Data are presented as the mean ± standard error of the mean (SEM). Experiments are considered significant at p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****). Individual data points represent biological replicates unless otherwise stated. Source data are provided as a Source data file.