Fig 2: Depletion of macrophages has limited effects on functional CSCs in Met-1 tumors from lean or obese mice. Lean or obese FVB/N female mice were transplanted with Met-1 tumor cells, and received treatment with either anti-F4/80 or IgG control antibodies (n = 3–4/group). Tumor growth curves (A) and end stage tumor masses (B) from treated mice. Tumor growth and end stage tumor masses analyzed with two-way ANOVA with Tukey’s multiple comparison test (*p < 0.0001 HFD IgG compared to HFD F4/80; **p < 0.0001 HFD IgG compared to CD IgG and HFD IgG compared to CD F4/80). (C) Isolated Met-1 tumor cells were plated in limiting dilution on non-adherent plates to form tumorspheres (n = 3–4/group). Tumorspheres were dissociated and re-plated to form secondary tumorspheres (two-way ANOVA with Tukey’s multiple comparison test). (D) Isolated Met-1 tumor cells were plated on collagen-coated migration chambers, and invading cells were quantified (n= 3/group; two-way ANOVA with Tukey’s multiple comparison test). (E) Quantification of Met-1 tumor cell invasive foci into surrounding adipose tissue (n = 3–4/group; two-way ANOVA with Tukey’s multiple comparison test). Bars represent mean ± s.e.m. Magnification bars = 50 µm.

Fig 3: Ductal deletion of Hnf1b leads to chronic pancreatitis at 2 months. (A) Pancreas weight and relative pancreas weight/body weight of 2-month-old mice (control, n = 17; mutant, n = 11). (B–F) H&E staining. Mutant pancreata show dramatic loss of (C) acinar tissue and (E and F) ADM. (G and H) Amylase (green) and Sox9 (red) immunostaining. (H’) Nuclear Sox9+ staining in amylase+ cells is characteristic of ADM in mutants. (I) RT-qPCR of acinar markers. (J) H&E staining shows lymphocyte infiltration and lipomatosis in mutants. (K) RT-qPCR of immune infiltrates (F4/80, CD2, CD19), cytokines (IL10), and chemokines (CCL2, CCL5, CXCL10). (L and M) Macrophage marker F4/80 (brown) immunostaining. (N and O) Adipocyte marker Fabp4 (green) immunostaining and (P) RT-qPCR of PPARγ. (Q and R) Masson’s trichrome staining. (S) RT-qPCR of Col1A1. (T and U) Vimentin (brown) immunostaining. (V) RT-qPCR of SMA, vimentin (Vim), N-Cad, and E-Cad. Scale bars: (B and C) 2 mm; (D–H, J, L, M, and Q–U) 50 μm; and (N and O) 100 μm. Nuclei are stained with 4′,6-diamidino-2-phenylindole (blue). Control, n ≥ 3; mutant, n ≥ 3 for immunostaining; and control, n = 8; mutant, n = 5 for RT-qPCR. **P < .01; ***P < .001. mRNA, messenger RNA.

Fig 4: Kupffer cells are the primary cell type phagocytizing desialylated platelets in the liver.A, B Representative confocal microscopic images. CD41 marks platelets, F4/80 marks Kupffer cells, CD31 labels sinusoidal endothelium, and hepatocytes were detected by autofluorescence. C Pseudocolored TEM micrographs of WT or Clec4f−/− ultrathin sections. Red arrow marks a phagocytized platelet. Blue arrows mark potential interactions between platelets and sinusoidal endothelium.

Fig 5: Monocytes, neutrophils, and macrophages increased in chronic CS-treatment emphysema model miceFlow cytometry was used to measure the percentage of CD11b+ Ly6C+ cells (‘inflammatory’ monocytes), CD11b+ Ly6G+ cells (neutrophils), and CD11b+ F4/80+ cells (macrophages) in the small intestine (A) and colon (B) of air-exposed control and emphysema model mice. (C) Immunohistochemical evaluation of F4/80 expression in the small intestine (upper) and colon (lower) from air-exposed control or emphysema model mice (n=5 per group). Scale bar = 100 μm. (D) The ratio of M1 (CD11b+F4/80+CD86+ cells) to M2 (CD11b+F4/80+CD206+ cells) were assessed in the small intestine and colon from three groups (n=5).