Fig 2: Alcohol-induced morphologic changes in microglia are partially restored by CCR2/5 inhibition. a Representative images of hippocampal microglia (CX3CR1-GFP+; green) displaying altered morphologies in pair-fed (PF) vs alcohol-fed (EtOH) mice, acquired at × 63 magnification. b Quantification of average hippocampal microglia soma area (pixel2) and c perimeter distance (pixel length) in mice fed control or alcohol diet or alcohol-fed mice treated with CCR2/5 inhibitor. d Expression of various activation markers were measured by flow cytometry in CD11b+ CD45lo microglia including CD86, CD68, MHC-II, CD163, and CD206. Data are mean ± SEM, n = 3 mice/group and 3–10 sections/mouse for a–c and n = 6–7 mice/group for d. *p < 0.05 by one-way ANOVA.

Fig 3: Long‐term exercise restores the foreign‐body phagocytic capacity of Kupffer cells (KCs) in p62‐KO mice. (a) Blood lactate concentration pre‐ and post‐exercise. The body mass of wild‐type mice fed a standard diet (WT, n = 3), resting p62‐KO mice fed a standard diet (p62‐KO‐Rest, n = 8), and p62‐KO fed a standard diet that exercised for 4 weeks (p62‐KO‐Ex, n = 8). (b) Latex bead‐phagocytic capacity of F4/80 positive cells (KCs) and their mean fluorescence intensity (MFI) in the livers of mice in each group. The MFI of the latex beads was determined using flow cytometry (n = 3/group). (c and d) Expression of the surface markers CD68 and toll‐like receptor (TLR) 4 (C), and CD11b (M1) and CD206 (M2) (d) in KCs from the livers of mice in each group (n = 3–4/group). (e) Lipopolysaccharide (LPS) concentrations in the peripheral and portal circulations and the differences between these (n = 5–6/group). (f) Dehydroepiandrosterone (DHEA) concentration in the peripheral blood (n = 6–8/group). Values are mean ± SE. *p < 0.05, compared with the WT group. † p < 0.05, compared with the p62‐KO‐Rest group

Fig 4: M2 macrophage polarization is reduced in SCC from OSM-KO mice and cSCC development is similar in WT and OSMR-KO mice(A) Tumor cells and infiltrating cells were isolated from OSM-KO mice and their WT littermates. Cells were stained with anti-CD45 V500, anti-CD11b AF488, anti-CD68 PerCPCy5.5, anti-CD206-BV421 or anti-iNOS-APC antibody and analyzed by flow cytometry. (B) Data are presented as the mean ± SEM of the percentage of M1 (iNOS+/CD206-) and M2 (iNOS-/CD206+) macrophages among CD45+ cells per tumor from four independent experiments, Two-way ANOVA test with Bonferroni post-tests, *p < 0.05. (C) OSMR-KO mice and their WT littermates were subcutaneously injected with 1.106 PDVC57 cells and tumor volume measured each week using a digital caliper. Data are presented as the mean ± SEM tumor volume (mm3) from two independent experiments, Two-way ANOVA test with Bonferroni post-tests.

Fig 5: RXRα deficiency impacts multiple RTM subsets.Analyses of RTM subsets from 8- 10-wk-old Rxrafl/fl (wild-type [WT]) and VavCre+Rxrafl/fl (knockout [KO]) mice. Flow cytometry gating strategies are shown in Supplemental Fig. 2. (A) Representative flow cytometry dot plots and graphs show frequency and cell numbers among CD45+ cells of splenic red pulp macrophages (RPMs) (CD11bloF4/80hiVCAM1+), (B) Upper row, Immunohistochemistry staining of IBA-1 (pan-macrophage marker) on splenic sections. White scale bars, 50 μm. Bottom row, Representative microscopy image of splenic sections showing iron deposits by Prussian blue stain. Black scale bars, 25 μm. (C) Representative flow cytometry dot plots and graphs show frequency and cell numbers among CD45+ cells of bone marrow erythroblastic island macrophages (EIMs) (CD11bloF4/80hiVCAM1+). (D) Representative flow cytometry dot plots and graphs show frequency and cell numbers among CD45+ cells of lung alveolar macrophages (AMs) (CD11c+Siglec-F+). (E) Immunohistochemistry staining of IBA-1 (pan-macrophage marker) on lung sections. Scale bars, 50 μm. (F–K) Representative flow cytometry dot plots and graphs show frequency and cell numbers among CD45+ cells of (F) liver Kupffer cells (KCs) (F4/80+Tim4+CD11b+CD68+), (G) large and small peritoneal cavity macrophages (LPMs, SPMs) (CD11b+F4/80hi and CD11b+F4/80lo), (H) small intestine macrophages (SI gMacs) (CD11b+CD64+CD11cloF4/80+) (I) large intestine macrophages (Colon gMacs) (CD11b+CD64+CD11cloF4/80+), (J) brain microglia (CD45loCD11b+), and (K) skin Langerhans cells (LCs) (CD11b+F4/80+). Data are representative of three independent experiments. Error bars are SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.