Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 9 Immunohistological comparison of CD163 and CD206 expression in various murine tumors. Immunohistochemical staining of CD163 and CD206 in MCA205, MC38, hepatoma, and histiocytic sarcoma. Scale bars: 200 μm.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Comparison, Expressing, Immunohistochemical staining, Staining

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 11 Expression of macrophage surface markers on recovered macrophages after clodronate liposome-mediated macrophage depletion. (A) CCL2 gene expression in the liver and spleen 3 days and 1 month post-clodronate administration in C57BL/6 mice. Data presented as mean ± SD. **, p < .01. (B) Western blot analyses of CD163, CD204, CD206 in the liver and spleen 1 month post-clodronate administration in C57BL/6 mice.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Expressing, Gene Expression, Western Blot

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 13 Expression of macrophage surface markers in the liver and spleen of obese mice. (A) Gene expression of CCL2, TNF-α, and IL-6 in the liver and spleen 3 months post-ND or HFD administration. Data presented as mean ± SD. *, p < .05. **, p < .01. (B) Western blot analyses of CD163, CD204, CD206 in the liver and spleen 3 months post-ND or HFD administration.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Expressing, Gene Expression, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: Adipose tissue inflammation in WT and OSMRβ−/− mice at 8 weeks on the HFD. The mice (8 weeks old) were fed an HFD for 8 weeks. A, total number of F4/80-positive cells per weights of adipose tissue in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice (n = 6). EWAT, epididymal white adipose tissue. B and C, the percentages of CD11c-positive cells (B) and CD206-positive cells (C) among the F4/80-positive cells in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice. D, the percentages of CD11c/CD206-double-negative cells among the F4/80-positive cells in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice. E and F, the mRNA expression levels of inflammatory and anti-inflammatory markers (TNF-α, IL-1β, IFN-γ, CCR2, MCP-1, TLR4, IL-6, MGL1, MGL2, IL-10, IL-13, and adiponectin) in the adipose tissue (E) and SVF (F) of WT, OSMRβ−/−, and OSMRβ−/−(PF) mice (n = 6). ADPN, adiponectin. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 WT versus OSMRβ−/− mice; #, p < 0.05 WT versus OSMRβ−/−(PF) mice, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: Adipose tissue inflammation and glucose metabolism in WT and OSMRβ−/− mice at 2 and 4 weeks on the HFD. A, total number of F4/80-positive cells per weights of adipose tissue in WT and OSMRβ−/− mice (n = 6). B and C, the percentages of CD11c-positive cells (B) and CD206-positive cells (C) among the F4/80-positive cells in WT and OSMRβ−/− mice. D, the percentages of CD11c/CD206-double-negative cells among the F4/80-positive cells in WT and OSMRβ−/− mice. E and F, the mRNA expression levels of inflammatory and anti-inflammatory markers (TNF-α, IL-1β, IFN-γ, CCR2, MCP-1, TLR4, IL-6, MGL1, MGL2, IL-10, IL-13, and adiponectin) in the adipose tissue (E) and SVF (F) of WT and OSMRβ−/− mice (n = 6). G–N, the results of the ipGTTs (G and K) and ITTs (I and M) in WT and OSMRβ−/− mice at 2 weeks (G–J) and 4 weeks (K–N) on the HFD (n = 6). The AUC for blood glucose on the ipGTTs (H and L) and ITTs (J and N) was shown. ADPN, adiponectin. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 WT versus OSMRβ−/− mice, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: The effects of OSM on glucose metabolism, adipose tissue inflammation, and hepatic steatosis in ob/ob mice. ob/ob mice were injected intraperitoneally with either vehicle or recombinant mouse OSM (12.5 ng/g of body weight) twice a day for 7 days. A, the body weights in vehicle- and OSM-treated ob/ob mice (n = 6). B, the tissue weights in vehicle- and OSM-treated ob/ob mice (n = 6). EWAT, epididymal white adipose tissue; SWAT, subcutaneous white adipose tissue. C and D, blood glucose (C) and serum insulin (D) levels in vehicle- and OSM-treated ob/ob mice in the fasted states (n = 6). In the fasted states, mice were fasted overnight before the experiments. E–H, the results of the ipGTTs (E) and ITTs (G) in vehicle- and OSM-treated ob/ob mice (n = 6). The AUC for blood glucose on the ipGTTs (F) and ITTs (H) is shown. For ipGTTs, mice were fasted for 16 h and intraperitoneally injected with d-glucose (0.5 g/kg of body weight). For ITTs, mice were fasted for 4 h and intraperitoneally injected with insulin (5 unit/kg of body weight). I, total number of F4/80-positive cells per weights of adipose tissue in vehicle- and OSM-treated ob/ob mice (n = 6). J and K, the percentages of CD11c-positive cells (J) and CD206-positive cells (K) among the F4/80-positive cells in vehicle- and OSM-treated ob/ob mice. L, the mRNA expression levels of anti-inflammatory markers (IL-10, IL-13, MGL1, and MGL2) in the adipose tissue of vehicle- and OSM-treated ob/ob mice (n = 6). M, Oil Red O and PAS staining of the livers of vehicle- and OSM-treated ob/ob mice. CV, central vein. Scale bar = 100 μm. N and O, the content of triglycerides (N) and total cholesterol (O) in the livers of vehicle- and OSM-treated ob/ob mice in the fed state (n = 6). In the fed state, mice were fasted for 4 h before the experiments to eliminate the feeding effects on lipid metabolism. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 vehicle versus OSM, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Injection, Recombinant, Expressing, Staining

Journal: Frontiers in Immunology

Article Title: S100A12 Expression Is Modulated During Monocyte Differentiation and Reflects Periodontitis Severity

doi: 10.3389/fimmu.2020.00086

Figure Lengend Snippet: S100A12 + monocyte-derived cells in gingival tissue from periodontitis patients and healthy controls. (A) Representative immunohistochemical staining of S100A12 in gingival tissue from a periodontitis patient and control. (B) Immunofluorescent staining of gingival tissue from periodontitis patient showing the presence of S100A12 (red) and CD206 (green). Nuclei was counterstained with DAPI (blue). (C) Western blot analysis of S100A12 monomer in gingival tissue from periodontitis patients ( n = 6) and controls ( n = 6), and S100A12 expression presented as normalized intensity to β-actin. (D) Representative zebra plot showing the gating strategy of monocyte-derived cells (MCs) in gingival tissue based on CD14 and CD64 expression on single, live, CD45 + cells, and zebra plots depicting the S100A12 + MCs in health and in periodontitis. Proportion of S100A12 + cells is included in the plot. (E) Percentage of CD14 + CD64 + cells within the CD45 + compartment in gingival tissue from periodontitis patients ( n = 6) and controls ( n = 6). (F) Percentage of S100A12 + cells within the CD14 + CD64 + compartment in gingival tissue from periodontitis patients ( n = 6) and controls ( n = 6). (G) Percentage of CD206 + cells in S100A12 + MCs in gingival tissue from periodontitis patients ( n = 6) and controls ( n = 6). (H) Spearmen correlation between the percentage of S100A12 + MCs and the percentage of CD206 + MCs. White and black circles indicate controls and periodontitis participants, respectively. Data are presented as mean ± SEM. Differences were calculated with Mann–Whitney test, * p < 0.05, ** p < 0.001.

Article Snippet: Double labeling was performed on gingival tissue using the following antibodies: mouse anti-S100A12 (clone OTI1D1) and rat anti-CD206 (clone 309210, R&D Systems, Minneapolis, MN, USA) or isotype controls.

Techniques: Derivative Assay, Immunohistochemical staining, Staining, Western Blot, Expressing, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

doi: 10.1016/j.xcrm.2024.101397

Figure Lengend Snippet: T cell infiltration into tumors occurs independent of the gut and tumor microbiome (A) Study design for (B)–(K). (B) 16S rRNA levels in tumor and stool from mice orthotopically injected with cold (69) tumor cells (n = 10) or hot (2838c3) tumor cells and treated with (n = 15) and without (n = 20) antibiotics. (C and D) Quantification of CD3 + (C) and CD8 + and FOXP3 + (D) T cells from hot tumors of mice treated with (n = 19) or without (n = 13) antibiotics. (E) 16S rRNA levels in tumor from mice orthotopically injected with cold (69) tumor cells and treated with (n = 5) and without (n = 5) antibiotics. (F) Quantification of CD8 + and FOXP3 + T cells from cold tumors of mice treated with (n = 5) and without (n = 5) antibiotics. (G) Number of DEGs. (H) Bar graph displaying overrepresentation analysis of DEGs in indicated gene sets. (I and J) Tumor weights at day 20. (K) Quantification of intra-tumoral CD19 + cells. (L and M) Mean fluorescence intensity (MFI) of MHC class II (L) and CD206 (M) on CD11b + F4/80 + intra-tumoral macrophages. Data were pooled from two to three experiments (B–D and J–M) or are representative of two independent experiments (E–I). Statistical significance was calculated using one-way ANOVA with Dunnett’s test (B, L, and M) and a two-tailed Mann-Whitney test (C–F and I–K). Data are represented as scatterplots (mean ± SD). NS, not significant.

Article Snippet: Rat monoclonal anti-mouse CD206 (MR6F3) , eBioscience , 17-2061-82; RRID:AB_2637420.

Techniques: Injection, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

doi: 10.1016/j.xcrm.2024.101397

Figure Lengend Snippet:

Article Snippet: Rat monoclonal anti-mouse CD206 (MR6F3) , eBioscience , 17-2061-82; RRID:AB_2637420.

Techniques: Recombinant, Sequencing, Software