Efficacy of CCL2 depletion and extent of intestinal inflammation in SLICK-H ^ER-Cre Ccl2 ^fl/fl mice. (A) Complementary to Fig. 5 D: confocal images of colonic myenteric plexus from LPS-injected Ccl2-RFP^fl/fl mice, stained with MHCII (MMs), tdTomato (Ccl2-RFP), and Hu (neuronal somas) antibodies. White arrowheads indicate CCL2⁺Hu⁺ neurons. Scale bars, 50 μm. (B) Confocal images of large bowel muscularis from the inflamed tissue cross-section of a CD patient described in Fig. 3, M–O, stained with CCL2 or isotype control IgG and Hu (neuronal somas) antibodies, and DAPI (nuclei). Scale bars, 50 μm. (C) Confocal images of colonic myenteric plexus from tamoxifen- and 1×DSS-treated SLICK-H^ER-CreCcl2-RFP^fl/fl (Cre⁺) and Ccl2-RFP^fl/fl (Cre^–) littermates at week 1, stained with MHCII (MMs), tdTomato (Ccl2-RFP), and Hu (neuronal somas) antibodies. White arrowheads indicate CCL2⁺ Hu⁺ neurons. Scale bars, 50 μm. (D) Complementary to Fig. 5 F: FC plots showing proportions of Ly6c⁺ and Ly6c^– Mo-MM subsets and their percentages in colons of tamoxifen- and 3×DSS- (2.15%) or water-treated SLICK-H^ER-CreCcl2-RFP^fl/fl (Cre⁺) and Ccl2-RFP^fl/fl (Cre^–) littermates. (E) Percentage of total viable cells (left) and absolute cell counts (right) of colonic M-PMNs in mice described in D, quantified by FC (DSS Cre^– and Cre⁺, n = 8 per group). Baseline shows the mean value for tamoxifen- and water-treated Cre^– littermates (water Cre^–, n = 5–6). (F) Percentage of total viable cells (left) and absolute cell counts (right) of colonic M-T cells in mice described in D, quantified by FC (DSS Cre^– Cre⁺, n = 8 per group; water Cre^– baseline, n = 5–6). P values above each column indicate statistical differences against baseline. (G) Percentage of body weight change in tamoxifen- and 3×DSS-treated mice described in D (DSS Cre^– and Cre⁺, n ≥ 10 per group). (H) Fecal lipocalin-2 in tamoxifen- and 3×DSS-treated mice described in D (DSS Cre^– and Cre⁺, n ≥ 10 per group). (I) Percentage of total viable cells (left) and absolute cell counts (right) of colonic mucosal Ly6g⁺ neutrophils in mice described in D, quantified by FC (DSS Cre^– and Cre⁺, n = 7–8 per group; water Cre^– baseline, n = 5–6). P values above each column indicate statistical differences against baseline. (J) Percentage of blood Ly6c^hi monocytes among total viable CD45⁺ blood cells in mice described in D, quantified by FC (DSS Cre^– and Cre⁺, n = 4 per group; Water Cre^– baseline, n = 3). All graphs show the mean ± SEM, n—mouse. Data are representative of at least two independent experiments with similar results. Data are pooled from 3 smaller experiments (G and H). Statistical analyses: unpaired Student’s t test (E, F, I, and J) and multiple t tests (G and H), *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001.