Fig. 2

Depletion of TREM-1 affects neutrophil and macrophage infiltration, inflammatory cytokines induction and decreases IL-22 production by ILC3 cells in colons upon DSS treatment. a-b Contour plot of FACS analysis data showed the gating strategy employed to identify neutrophils, DCs, CD4 T cells, CD8 T cells and ILC cells in colons from water- or DSS-treated WT and TREM-1 KO mice (n = 10/group) on day 7 post-DSS initiation. Lamina propria macrophages were subcategorized into CD45⁺CD11b⁺CD11c⁻CD64⁺ and further presented by histogram of MHCII staining. c FACS analysis of the percentage and absolute number of the indicated immune cells among colonic cells from the indicated mice in (a-b). Data are the mean ± SEM and representative of at least three independent experiments. d Real-time PCR analysis of mRNA levels of the indicated genes in colonic tissue of DSS-treated WT or TREM-1 KO mice on day 9. Data are presented as fold change relative to the GAPDH mRNA level and are representative of at least 3 independent experiments involving 20 mice per group. e Contour plot of FACS analysis data showed the gating strategy employed to identify distinct ILC subsets by intracellular staining with specific transcription factors. f FACS analysis of the frequency and absolute number of the indicated ILC subsets among live CD45⁺LIN⁻TCRβ⁻NK1.1⁻Thy1.2⁺ cells isolated from colons of water-treated WT and TREM-1 KO mice and DSS-treated WT and TREM-1 KO mice (n = 8/group) on day 7 post-DSS initiation. g The percentage and absolute number of CD45⁺LIN⁻TCRβ⁻NK1.1⁻Thy1.2⁺RORγt⁺IL-22⁺ ILC3 cells and CD45⁺TCRβ⁺IL-22⁺ T cells among colonic cells from the indicated mice (n = 10/ group) on day 7 post-DSS initiation. Data are the mean ± SEM and are representative of 3 independent experiments. h Quantitation of IL-22 as determined by ELISA in 0.1 g of colonic tissue from water-treated WT and TREM-1 KO mice (n = 5/group) and DSS-treated WT and TREM-1 KO mice (n = 5/group) on day 7 post-DSS initiation. Data are mean ± SEM and representative of two independent experiments. Statistical significance was determined by Mann-Whitney nonparametric test. * p < 0.05 or **, p < 0.01as indicated