Journal: Journal of Animal Science and Biotechnology

Article Title: Heat stress induced testicular impairment is related to orchitis and complement activation in Rongchang boars

doi: 10.1186/s40104-025-01296-5

Figure Lengend Snippet: Testicular immunity and orchitis under HS conditions. A and B GO (A) and KEGG (B) enrichments based on Gene Set Enrichment analysis. C Protein-protein interaction network. D Representative photos of collagen fibers stained by Masson in the testes. E and F Levels of TNF-α (E) and IL-1β (F) in plasma isolated from the spermatic vein. G Representative flow cytometry quantifications of macrophages in the testes. H and I Percentages of CD68 + CD163 + and CD68 + cells among CD45 + cells in the testes. * P < 0.05, ** P < 0.01

Article Snippet: The remaining cells were resuspended in 100 μL Stain Buffer (BD, Franklin Lakes, New Jersey, USA) containing mouse anti-pig CD45: FITC (Bio-Rad, Hercules, California, USA), mouse anti-pig CD163: RPE (Bio-Rad, Hercules, California, USA), and BV421 rat anti-CD11b (BD, Franklin Lakes, New Jersey, USA) at 4 °C for 30 min. Each suspension was centrifuged at 350 × g for 5 min at 4 °C to remove the supernatant.

Techniques: Staining, Clinical Proteomics, Isolation, Flow Cytometry

Journal: Journal of Animal Science and Biotechnology

Article Title: Heat stress induced testicular impairment is related to orchitis and complement activation in Rongchang boars

doi: 10.1186/s40104-025-01296-5

Figure Lengend Snippet: Changes in the TM profile and complement pathway under HS conditions. A tSNE graph of identified cells. B Percentages of cells in each group. C KEGG enrichment of the DEGs from TMs. D Expression levels of C1QA , C1QB , and C1QC in testicular cells as assessed by scRNA-seq. E tSNE graph of TM subclusters in each group. F Percentages of subclusters in each group. G KEGG enrichment of marker genes in Subcluster 5. H Expression levels of C1QA , C1QB , and C1QC in TM subclusters as assessed by scRNA-seq. I KEGG enrichment of DEGs from TM Subcluster 5. J Addmodulescore of subclusters. K tSNE graph of C1QA , C1QB , C1QC , and CD163 as assessed by scRNA-seq

Article Snippet: The remaining cells were resuspended in 100 μL Stain Buffer (BD, Franklin Lakes, New Jersey, USA) containing mouse anti-pig CD45: FITC (Bio-Rad, Hercules, California, USA), mouse anti-pig CD163: RPE (Bio-Rad, Hercules, California, USA), and BV421 rat anti-CD11b (BD, Franklin Lakes, New Jersey, USA) at 4 °C for 30 min. Each suspension was centrifuged at 350 × g for 5 min at 4 °C to remove the supernatant.

Techniques: Expressing, Marker

Journal: The FASEB Journal

Article Title: Proangiogenic effects of tumor cells on endothelial progenitor cells vary with tumor type in an in vitro and in vivo rat model

doi: 10.1096/fj.201800135rr

Figure Lengend Snippet: Figure 8. Immunohistochemical staining for tumor associated macrophages and tumor cells in tumor xenografts encapsulated in alginate/ fibrin hydrogels. A) Representative images of CD68, CD163, and cytokeratin immunohisto- chemical staining in CC531 and MCR86 groups. B) Representative images of CD68, CD163, and vimentin immunohistochemical staining in ROS-1 group. Scale bars, 100 mm.

Article Snippet: To whether investigate macrophages infiltrated the implanted hydrogels, an anti-rat CD68 mouse monoclonal antibody (1:300, clone ED1) and CD163 (1:200, clone ED2; both from Bio-Rad) were used after pretreatment with pronase (MilliporeSigma).

Techniques: Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis

doi: 10.1038/s41598-021-93839-9

Figure Lengend Snippet: Analysis of tracheal wash derived macrophages. Flow cytometry results showing cross-reactivity of mouse anti-human CD163 antibody against equine tracheal macrophages. (a) Isotype control, (b) CD163 stained cells (c) overlay of CD163+ population on top of total cells. (d) Leishman stained cytospin preparations of CD163+ cells by light microscopy (× 20, scale bar = 50 μm). Data and image analysis was performed in FlowJo ® v10.5.3 https://www.flowjo.com/ .

Article Snippet: Briefly, the cells were first stained with a R-Phycoerythrin (PE)-conjugated CD163 primary antibody; subsequently, the cells were magnetically labelled with anti-PE MicroBeads (Miltenyi Biotec, cat no 130-105-639) before the cell suspension was loaded on a MACS ® LS Column (Miltenyi Biotec Ltd., cat n0 130-042-401), placed in the magnetic field of a MACS Separator.

Techniques: Derivative Assay, Flow Cytometry, Control, Staining, Light Microscopy

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet: Combination of CITE-seq, scRNA-seq, snRNA-seq, and spatial analyses enables generation of a human liver atlas and identification of bona fide human KCs, related to Figure 4 (A and B) Top DEGs (A) and DEPs (B) for the cell types from Figure 4 B. (C) Distinct profiles of cells or nuclei within the UMAP depending on isolation protocol used; 152,535 cells from ex vivo digestions and 15,063 nuclei. (D) Proportion of each cell type per patient profiled. (E) Proportion of indicated cell types as a % of total CD45 + cells calculated from ex vivo digested samples per surgery type. Ch; cholecystectomy, Re; resection, GB; gastric bypass. ∗ p < 0.05; one-way ANOVA with Bonferroni post-test. (F) Mapping of Visium UMAP zonation patterns onto tissue sections from patient H35 and H37. (G) Expression of indicated zonation genes in patients H35–H38 assessed by Molecular Cartography. (H and I) Expression of indicated proteins by MICS 100-plex protein analysis in the healthy (H) and steatotic (I) human liver. (J) Murine myeloid cells (cDC1s, cDC2s, Mig. cDCs, Macs, monocytes, and monocyte-derived cells; 42,922 cells) from mice fed the SD or WD for 24 or 36 weeks were isolated from Figure S5 J and re-clustered with TotalVI. (K) Distribution of cells in UMAP originating from SD- (purple) or WD- (yellow) fed mice. (L) Proportion of indicated cell types arising from mice fed the SD (purple) or WD (yellow). (M and N) Flow cytometry analysis of indicated cell populations in SD and WD-fed mice (24 weeks). Representative gating strategies (M) and absolute number of indicated populations (N). ∗ p < 0.05, ∗∗ p < 0.01 Student’s t test. Data are from 2 independent experiments with n = 5–6 per diet. (O and P) Top DEGs (O) and DEPs (P) for cell types from Figure 4 H. (Q) Top 25 Murine KC genes as expressed by the human myeloid cell clusters. (R) Mapping of KC signature onto Visium trajectory for healthy (purple) and steatotic (orange) livers. (S) Expression of VSIG4 mRNA within human myeloid cells. (T) Expression of VSIG4 (red) and CD163 (gray, top) or CD169 (gray, bottom) by MICS analysis in healthy human liver. (U) Representative images showing KC location (red) as assessed by MICS analysis in the healthy (left) and steatotic (right) human liver. PV, portal vein; CV, central Vein, dashed line indicates zones of steatosis. (V) Representative image of CD68 and CD163 staining in 10–15-year-old human liver paraffin sections. Image is representative of 6 different patients. (W) In silico gating strategy to isolate distinct myeloid cell populations identified from CITE-seq data. (X) Expression of VSIG4 and FOLR2 by live CD45 + cells also expressing CD14 in indicated human liver biopsies by flow cytometry. Data are representative of 21 biopsy samples analyzed.

Article Snippet: Anti-Human CD163 (REA406) PE , Miltenyi Biotec , 130-121-316; RRID: AB_2857545.

Techniques: Isolation, Ex Vivo, Expressing, Derivative Assay, Flow Cytometry, Staining, In Silico

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Human CD163 (REA406) PE , Miltenyi Biotec , 130-121-316; RRID: AB_2857545.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: CIBERSORT analysis of immune cell infiltration in ESCC samples from TCGA database. A CIBERSORT analysis showed immune cell infiltration and immune cell correlation. B There was macrophage infiltration in ESCC tumor tissues. C TIMER2.0 was used to analyze the correlation between ETV1 and M0 and M2 macrophage infiltration. D Correlation between ETV1 and M2-labeled CD206 or CD163 expression based on the GEPIA database

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Labeling, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: ETV1 expression was significantly elevated in tumor tissues of patients with ESCC. A Results of TCGA and GEPIA databases showed that ETV1 expression was markedly increased in tumor tissues of patients with ESCC. B Real time PCR was used to detect the mRNA expression of ETV1 in fresh frozen ESCC and adjacent samples. C Western blot was employed to determine the protein expression of ETV1 in ESCC and adjacent samples. D The expression of ETV1 in pathological progression of ESCC was tested by immunohistochemical staining. Scale bar, 100 µm. (E–F) The expression of ETV1 and CD206 or CD163 in ESCC tissues was examined by immunofluorescence double staining. Scale bar, 100 µm. The correlation between the fluorescence intensity of ETV1 and the number of CD206 + or CD163 + cells was analyzed. Data are mean ± SD. ****, p < 0.0001

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Double Staining, Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells

doi: 10.1007/s00262-024-03914-z

Figure Lengend Snippet: ETV1 facilitates M2 macrophage chemotaxis and polarization. A THP-1-M0 cells were placed in the upper chamber of transwell, and ESCC cells were placed in the lower chamber of transwell. After 24 h of co-culture, the cells migrated to the submembrane surface were detected by crystal violet staining. Scale bar, 100 µm. B , C ESCC cells were placed in the upper chamber of transwell, and THP-1-M0 cells were placed in the lower chamber of transwell. After 24 h of co-culture, the proportions of CD68 + CD206 + and CD68 + CD163 + in THP-1-M0 cells were tested by flow cytometry. Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001

Article Snippet: The following antibodies were used: human CD68-FITC (65,202, Proteintech, USA); human CD206-PE (65,155, Proteintech); human CD163-PE (65,169, Proteintech); mouse CD206 (12–2061-80, Invitrogen, USA) and mouse CD68 (137,005, Biolegend, USA).

Techniques: Chemotaxis Assay, Co-Culture Assay, Staining, Flow Cytometry

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Overview of flow cytometry analyses. (A) Gating strategy for identification of monocytes and macrophages. The peripheral blood is shown. Designation of population shown within each dot-plot is indicated above the dot-plot. Leukocytes were identified as viable (a) non-doublet (b) cells with typical light scatter properties of leukocytes (c) . Then, macrophages were gated simply as CD203a hi leukocytes (d) and marked with blue color. Monocytes were gated as CD203a low/- SWC8 - (e) CD172a hi (f) leukocytes where the CD203a low/- region was defined as the complementary region to the CD203a hi region. Then, SLA-DR + monocytes were marked with red color and SLA-DR - monocytes were marked with green color (g) . SLA-DR - region was defined as the complementary region to the SLA-DR + region. Gating order is shown in the scheme (h) . (B) Representative CD163 vs. CD14 dot-plots of macrophages (blue) and monocyte subpopulations (green: SLA-DR – , red: SLA-DR + ) in various body compartments of control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Flow Cytometry, Infection

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Primers for chemokines, chemokine receptors, CD62L and reference gene used in quantitative real-time PCR.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques:

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Monocyte subpopulations in various body compartments from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques:

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Representative pictures of immunohistochemical detection of CD163 + cells in tracheobronchial lymph node and spleen. CD163 + cells were detected in tracheobronchial lymph nodes from control (A) and APP-infected pigs (B) and in spleen from control (C) and APP-infected pigs (D) . Immunohistochemical visualization: horseradish peroxidase, brown substrate, hematoxylin counterstain; c, cortex; ca, central artery; e, ellipsoid; f, follicle; mz, marginal zone; pals, periarterial lymphatic sheath, rp, red pulp; s, subcapsular sinus.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Immunohistochemical staining, Infection

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Intensity of CD163, CD14 and SLA-DR expression by CD14 + /CD163 + monocytes in various body compartments. Data are shown as MFI (Median of fluorescence intensity) ± S.E.M. BM, bone marrow ( n = 5); PB, peripheral blood ( n = 5), UA, lungs-unaffected area ( n = 5), DZ, lungs-demarcation zone ( n = 4), NA, lungs-necrotic area ( n = 5), TBLN, tracheobronchial lymph node ( n = 5); MLN, mesenteric lymph node ( n = 4), spleen ( n = 4). The significant differences (Kruskal-Wallis test) amongst particular compartments are indicated.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing, Fluorescence

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Chemokine receptor, CD62L, CD163 and TBP1 expression by bone marrow and peripheral blood CD163 + monocytes from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Chemokine receptor, CD62L and CD163 expression in lungs from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Scheme of chemokines and corresponding chemokine receptor mRNA expression in various organs and CD163 + monocytes. Chemokines in bone marrow, necrotic area of the lungs and tracheobronchial lymph nodes and chemokine receptors in necrotic areas of the lungs and tracheobronchial lymph nodes which were up-regulated in the APP-infected pigs compared to control are highlighted in bold. Chemokine receptors in bone marrow and peripheral blood CD163 + monocytes which were expressed at relatively lower levels are highlighted in bold. Those expressed in relatively high levels are highlighted in bold and underlined. Arrangement of the scheme was created based on the review of Bonecchi et al. .

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing, Infection

Journal: Immunity

Article Title: Airway Epithelial Cell-Derived Colony Stimulating Factor-1 Promotes Allergen Sensitization

doi: 10.1016/j.immuni.2018.06.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-human CD163 , Fluidigm , GHI/64, cat#3154007B.

Techniques: Antibody Labeling, Virus, Recombinant, Control, Enzyme-linked Immunosorbent Assay, Software