Fig 2: Isolation of mouse liver KCs(A) Exposure of the portal vein and inferior vena cava after surgical dissection.(B) Perfusion with digestion buffer by inserting into the portal vein and flowing out from the inferior vena cava.(C and D) Liver turns light brown post thorough perfusion. (E) Liver homogenate post thorough mincing.(F) FACS gating strategy and the purity of sorted KCs which are CD31-/CD45+/CD11b+/F4/80+/Tim-4+.See also Methods video S1.

Fig 3: Monocyte-derived peritoneal macrophages from Nlrp3tm1Flv mice overexpress TIM4.a Representative flow cytometry plots showing identification of TIM4− and TIM4+ macrophages (F4/80+ CD11b+) in C57B6/J and Nlrp3−/− (Nlrp3tm1Flv) mouse peritoneum. Cells were pre-gated on single, live and CD45+ populations. b Bar plot of the total number of macrophages (CD11b+ F4/80+) in each peritoneum of each mouse (Nlrp3−/−, n = 4; C57B6/J, n = 4). P value was calculated using an unpaired t-test. Error bars represent standard deviation. c Representative histogram of distribution of TIM4− and TIM4+ macrophages (F4/80+ CD11b+) in C57B6/J and Nlrp3−/− (Nlrp3tm1Flv) mouse peritoneum. d Bar plot of TIM4− and TIM4+ macrophages (F4/80+ CD11b+) in C57B6/J, Nlrp3−/− (Nlrp3tm1Flv) and Casp1/11−/− mouse peritoneum. Percentages above bars show the mean value for each group. (Nlrp3−/−, n = 7; C57B6/J, n = 7; Casp1/11−/−, n = 8). Adj. P values were calculated using a two-way ANOVA with Turkey’s multiple comparison testing. *adj. P < 0.05, **adj. P < 0.01, ***adj. P < 0.001, ****adj. P < 0.0001, ns not significant (adj. P > 0.05). Error bars represent standard deviation. e Bar plot of TIM4− and TIM4+ macrophages (F4/80+ CD11b+) in C57B6/J and 129S2 mouse peritoneum. Percentages above bars show the mean value for each group (C57B6/J, n = 4; 129S2, n = 4). P values were calculated using multiple unpaired parametric t-tests. FDR (q) was calculated using Benjamini–Hochberg correction. *q < 0.05, **q < 0.01, ***q < 0.001, ****q < 0.0001, ns not significant (q > 0.05). Error bars represent standard deviation. f log2 normalised counts of Timd4 from RNAseq of Nlrp3−/− and Nlrp3+/+ (Nlrp3tm1Flv) BMDMs at baseline and after 3 and 24 h of LPS (10 ng/ml) stimulation (Nlrp3−/−, n = 4; Nlrp3+/+, n = 4). *adj. P < 0.05, **adj. P < 0.01, ***adj. P < 0.001, ****adj. P < 0.0001, ns not significant. Wald Test, Benjamini–Hochberg corrected.