anti ccr2  (Proteintech)

Journal: PLOS One

Article Title: Single-cell transcriptomic analysis reveals the association of Ccl6 + Ccr2 + Arg1 + macrophages with renal interstitial fibrosis in AKI

doi: 10.1371/journal.pone.0332026

Figure Lengend Snippet: (a) Analysis of cell-cell communication shows differences in interaction number and weight among cell subtypes in AKI and sham control groups. In the AKI group, macrophages exhibit increased interactions with themselves, neutrophils, and lymphocytes. (b) Ligand-receptor interaction weights indicate enhanced signal reception and outgoing signaling by macrophages in AKI kidneys compared to controls. (c) Intercellular signaling pathway analysis reveals intensified communication among immune cells, particularly within macrophage subtypes, in the AKI group. (d) Chemokine CCL signaling pathways are significantly upregulated in AKI macrophages, indicating activation of this pathway. (e) Ccl6/Ccr2 as the key driver of macrophage-macrophage interactions, promoting the recruitment of bone marrow-derived macrophages to the injured kidney and potentially influencing renal repair. (f) Validation of CCL signaling pathway activation in AKI. CellChat analyses across all AKI datasets at 7 days post-injury revealed consistent activation of the CCL signaling pathway, with the Ccl6–Ccr2 demonstrating strong interactions and emerging as a significantly enriched communication axis.

Article Snippet: The primary antibodies used were Ccl6 (Bioss, Beijing Biosynthesis Biotechnology Co., Ltd. bs-2476R, using at a 1:100 dilution), Ccr2 (Proteintech Group, Inc., 16153–1-AP, using at a 1:100 dilution), and Arg1 (Proteintech Group, Inc., 66129–1-Ig, using at a 1:2000 dilution).

Techniques: Control, Protein-Protein interactions, Activation Assay, Derivative Assay, Biomarker Discovery

Journal: PLOS One

Article Title: Single-cell transcriptomic analysis reveals the association of Ccl6 + Ccr2 + Arg1 + macrophages with renal interstitial fibrosis in AKI

doi: 10.1371/journal.pone.0332026

Figure Lengend Snippet: (a) UMAP visualization of 5,009 macrophages from Control and AKI mouse kidney tissues, reclustered into six distinct subpopulations. (b-c) Heatmap and FeaturePlot showing the marker genes for each macrophage subpopulation. (d) Pseudotime analysis using Monocle 3 reveals a differentiation trajectory from Stage I to Stage II. (e) Heatmap of pseudotime-ordered gene expression, highlighting differentially expressed genes along the trajectory. Ccl6 and Ccr2 showed significant expression at the terminal stage of the trajectory. (f) FeaturePlot showing the expression patterns of Ccl6 and Ccr2 across macrophage clusters. A subset of Cluster 0 co-expresses both genes, suggesting a potential autocrine loop, while Cluster 3 predominantly expresses Ccl6 alone, supporting a paracrine model.

Article Snippet: The primary antibodies used were Ccl6 (Bioss, Beijing Biosynthesis Biotechnology Co., Ltd. bs-2476R, using at a 1:100 dilution), Ccr2 (Proteintech Group, Inc., 16153–1-AP, using at a 1:100 dilution), and Arg1 (Proteintech Group, Inc., 66129–1-Ig, using at a 1:2000 dilution).

Techniques: Control, Marker, Gene Expression, Expressing

Journal: PLOS One

Article Title: Single-cell transcriptomic analysis reveals the association of Ccl6 + Ccr2 + Arg1 + macrophages with renal interstitial fibrosis in AKI

doi: 10.1371/journal.pone.0332026

Figure Lengend Snippet: (a) Differential expression analysis of macrophages in the AKI group compared to the Control group. A total of 132 genes were upregulated, including Ccl6 and Ccr2 , while 295 genes were downregulated. (b) The FeaturePlot displays the top 10 upregulated genes in macrophages from AKI kidneys. (c) UMAP visualization and violin plots showing the expression distribution of Ccl6 and Ccr2 across cell clusters in the AKI group. (d) Correlation analysis reveals positive relationships between Ccl6 , Ccr2 , and key genes such as Retnla , Fn1 , and Arg1 . (e) GO enrichment analysis of 132 upregulated genes in macrophages from the AKI group. The genes are enriched in pathways such as cytokine-mediated signaling, leukocyte migration, and myeloid leukocyte chemotaxis. (f) Heatmap of co-expressed genes involved in the enriched pathways. (g) Gene relationship network showing the co-expression of Ccl6 and Ccr2 in pathways related to cytokine-mediated signaling, chemokine activity, and leukocyte migration. (h) Protein-Protein Interaction (PPI) network of upregulated genes in the AKI group. Ccl6 and Ccr2 demonstrate strong interactions within three core pathways: chemokine receptor-ligand binding, chemokine-mediated signaling, and cellular responses to chemokines. (i) Ccl6 + Ccr2 + Arg1 + macrophages exhibit high Spp1 , Lgals3 , and Mmp12 expression, suggesting pro-fibrotic remodeling potential.

Article Snippet: The primary antibodies used were Ccl6 (Bioss, Beijing Biosynthesis Biotechnology Co., Ltd. bs-2476R, using at a 1:100 dilution), Ccr2 (Proteintech Group, Inc., 16153–1-AP, using at a 1:100 dilution), and Arg1 (Proteintech Group, Inc., 66129–1-Ig, using at a 1:2000 dilution).

Techniques: Quantitative Proteomics, Control, Expressing, Migration, Chemotaxis Assay, Activity Assay, Ligand Binding Assay

Journal: PLOS One

Article Title: Single-cell transcriptomic analysis reveals the association of Ccl6 + Ccr2 + Arg1 + macrophages with renal interstitial fibrosis in AKI

doi: 10.1371/journal.pone.0332026

Figure Lengend Snippet: (a) Schematic representation of the experimental workflow using the uIRI model to induce AKI. Kidney samples were harvested 7 days post-injury (n = 6). (b) Transcriptional analysis of Ccl6 , Ccr2 , Retnla , and Arg1 expression levels in kidney tissue, showing significant upregulation in the AKI group compared to sham controls (n = 6). (c) Histological analysis at day 7 post-AKI. H&E staining reveals tubular necrosis, epithelial detachment, and brush border loss. PSR and Masson staining demonstrate significant interstitial fibrosis. Quantification of tubular injury scores and fibrosis areas highlights incomplete renal repair and fibrotic remodeling (n = 6). Scale bar: 50 μm. (d, e) Colocalization analysis of Ccl6, Ccr2, and Arg1 in the OSOM region of the kidney at day 7 post-AKI. Immunofluorescence-based quantification showed significantly increased Pearson’s correlation coefficients among the three markers, indicating enhanced co-expression within the same cells (n = 6). Scale bar: 50 μm (f) Quantification of Ccl6 + Ccr2 + and Ccl6 + Ccr2 + Arg1 + cells revealed a significant increase in kidneys 7 days after AKI compared to sham controls (n = 6). (g, h) The infiltration of Ccl6 + Ccr2 + Arg1 + cells positively correlate with the severity of renal interstitial fibrosis. (i) Western blot analysis of Ccr2 protein levels in kidney tissue shows significant difference between AKI and sham controls. Results are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, ns : no significance.

Article Snippet: The primary antibodies used were Ccl6 (Bioss, Beijing Biosynthesis Biotechnology Co., Ltd. bs-2476R, using at a 1:100 dilution), Ccr2 (Proteintech Group, Inc., 16153–1-AP, using at a 1:100 dilution), and Arg1 (Proteintech Group, Inc., 66129–1-Ig, using at a 1:2000 dilution).

Techniques: Expressing, Staining, Immunofluorescence, Western Blot

Journal: PLOS One

Article Title: Single-cell transcriptomic analysis reveals the association of Ccl6 + Ccr2 + Arg1 + macrophages with renal interstitial fibrosis in AKI

doi: 10.1371/journal.pone.0332026

Figure Lengend Snippet: (a) Co-immunofluorescent staining for Arg1 and F4/80 in kidney tissues 7 days post-AKI showed a significant increase in expression compared to the Sham group, which was reduced following Ccr2 antagonist treatment (n = 3). Scale bar: 50 μm. (b, c) Co-localization analysis of Arg1 and F4/80 signals revealed a significant correlation, with no significant differences in Pearson’s correlation coefficient or Manders’ overlap coefficient among groups. (d) Quantification of Arg1 mean fluorescence intensity, Arg1-positive area percentage, and Arg1 + F4/80 + double-positive macrophages showed significant increases in the AKI group, which were reduced following Ccr2 blockade. (e, f) Masson and Sirius Red staining demonstrated a marked reduction in renal fibrosis upon Ccr2 inhibition (n = 3). Scale bar: 50 μm. (g) Confirmation of macrophage identity by Cd68 immunofluorescence staining in isolated bone BMDMs. Scale bar: 10 μm (h) CCK8 assay revealed no significant effect of 24-hour Ccl6 treatment on BMDM proliferation (n = 6). (i, j) Transwell migration assays showed a significant increase in BMDMs migration in the Ccl6-treated group (n = 6). (k, l) Flow cytometry analysis indicated an elevated mean fluorescence intensity of Arg1, confirming that Ccl6 promoted M2 polarization of BMDMs (n = 3). Results are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, ns : no significance.

Article Snippet: The primary antibodies used were Ccl6 (Bioss, Beijing Biosynthesis Biotechnology Co., Ltd. bs-2476R, using at a 1:100 dilution), Ccr2 (Proteintech Group, Inc., 16153–1-AP, using at a 1:100 dilution), and Arg1 (Proteintech Group, Inc., 66129–1-Ig, using at a 1:2000 dilution).

Techniques: Staining, Expressing, Fluorescence, Inhibition, Immunofluorescence, Isolation, CCK-8 Assay, Migration, Flow Cytometry

Journal: Frontiers in Endocrinology

Article Title: Protective effects of Shenkang injection against diabetic kidney disease via p38 MAPK/NFκB/MCP-1/CCR2 pathway inhibition

doi: 10.3389/fendo.2025.1596000

Figure Lengend Snippet: Protein and mRNA expression of CCR2 and MCP-1 in kidney tissues of mice in each group. (A–D) Expression and quantification of CCR2 and MCP-1 IHC. (E) WB expression of CCR2 and MCP-1. (F) Quantification of WB results for CCR2. (G) Quantification of WB results for MCP-1. (H) Quantification of CCR2 mRNA expression. (I) Quantification of MCP-1 mRNA expression. **P <0.01, compared with the model group. # P <0.05, ## P <0.01, compared with the control group. Data were analyzed by one way ANOVA (Mean ± SEM). IHC: N = 6. WB and RT-PCR: N=3. The x-axis represents different groups.

Article Snippet: Other reagents included: dagliflozin (NH3205); sodium pentobarbital (Beijing Chemical Reagent Company, catalog number: 020402); electron microscope fixative (Servicebio, catalog number: G1102); primary antibodies for immunohistochemistry and western blotting (WB): p-p38 (ImmunoWay, catalog number: YP0338), p38 (CST, catalog number: # 4511), MCP-1 (Thermo Fisher, catalog number: MA5-17040), CC motif chemokine receptor 2 (CCR2) (Proteintech, catalog number: 16153-1-AP), NFκB (CST, catalog number: 8242S), p-NFκB (ABways, catalog number: F107172), β-actin (CST, catalog number: # 4970); secondary antibodies: goat anti-rabbit (Abcam, catalog number: ab672), goat anti-rat (Abcam, catalog number: ab67891); hematoxylin (Solaibao, catalog number: G1080); eosin (Solaibao, catalog number: G1100); a Mallory Three-color Staining Kit (Solaibao, catalog number: G1355); a Glycogen Staining Solution (Beijing Legen, catalog number: DG0005); RIPA tissue/cell lysate (Lablead, catalog number: R1091); a BCA Protein Quantification Kit (Lablead, catalog number: B5001); a Protease Inhibitor Cocktail (Lablead, catalog number: C0101); and a Phosphatase Inhibitor Cocktail (Lablead, catalog number: R100).

Techniques: Expressing, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Endocrinology

Article Title: Protective effects of Shenkang injection against diabetic kidney disease via p38 MAPK/NFκB/MCP-1/CCR2 pathway inhibition

doi: 10.3389/fendo.2025.1596000

Figure Lengend Snippet: Cell viability and protein expression of CCR2 and MCP-1 in each group. (A) Histone expression as a loading control. (B) Western blot (WB) quantification of CCR2 protein expression. (C) WB quantification of MCP-1 protein expression. (D) mRNA expression levels of CCR2. (E) mRNA expression levels of MCP-1. (F) Cell viability in each group. *P <0.05, **P <0.01, compared with the model group. Data were analyzed by one way ANOVA (Mean ± SEM). CCK8: N = 6. WB: N=3. The x-axis represents different groups.

Article Snippet: Other reagents included: dagliflozin (NH3205); sodium pentobarbital (Beijing Chemical Reagent Company, catalog number: 020402); electron microscope fixative (Servicebio, catalog number: G1102); primary antibodies for immunohistochemistry and western blotting (WB): p-p38 (ImmunoWay, catalog number: YP0338), p38 (CST, catalog number: # 4511), MCP-1 (Thermo Fisher, catalog number: MA5-17040), CC motif chemokine receptor 2 (CCR2) (Proteintech, catalog number: 16153-1-AP), NFκB (CST, catalog number: 8242S), p-NFκB (ABways, catalog number: F107172), β-actin (CST, catalog number: # 4970); secondary antibodies: goat anti-rabbit (Abcam, catalog number: ab672), goat anti-rat (Abcam, catalog number: ab67891); hematoxylin (Solaibao, catalog number: G1080); eosin (Solaibao, catalog number: G1100); a Mallory Three-color Staining Kit (Solaibao, catalog number: G1355); a Glycogen Staining Solution (Beijing Legen, catalog number: DG0005); RIPA tissue/cell lysate (Lablead, catalog number: R1091); a BCA Protein Quantification Kit (Lablead, catalog number: B5001); a Protease Inhibitor Cocktail (Lablead, catalog number: C0101); and a Phosphatase Inhibitor Cocktail (Lablead, catalog number: R100).

Techniques: Expressing, Control, Western Blot

Journal: Frontiers in Endocrinology

Article Title: Protective effects of Shenkang injection against diabetic kidney disease via p38 MAPK/NFκB/MCP-1/CCR2 pathway inhibition

doi: 10.3389/fendo.2025.1596000

Figure Lengend Snippet: Schematic model of the effect of SKI on p38 MAPK/NF-κB/MCP1/CCR2 signaling pathway.

Article Snippet: Other reagents included: dagliflozin (NH3205); sodium pentobarbital (Beijing Chemical Reagent Company, catalog number: 020402); electron microscope fixative (Servicebio, catalog number: G1102); primary antibodies for immunohistochemistry and western blotting (WB): p-p38 (ImmunoWay, catalog number: YP0338), p38 (CST, catalog number: # 4511), MCP-1 (Thermo Fisher, catalog number: MA5-17040), CC motif chemokine receptor 2 (CCR2) (Proteintech, catalog number: 16153-1-AP), NFκB (CST, catalog number: 8242S), p-NFκB (ABways, catalog number: F107172), β-actin (CST, catalog number: # 4970); secondary antibodies: goat anti-rabbit (Abcam, catalog number: ab672), goat anti-rat (Abcam, catalog number: ab67891); hematoxylin (Solaibao, catalog number: G1080); eosin (Solaibao, catalog number: G1100); a Mallory Three-color Staining Kit (Solaibao, catalog number: G1355); a Glycogen Staining Solution (Beijing Legen, catalog number: DG0005); RIPA tissue/cell lysate (Lablead, catalog number: R1091); a BCA Protein Quantification Kit (Lablead, catalog number: B5001); a Protease Inhibitor Cocktail (Lablead, catalog number: C0101); and a Phosphatase Inhibitor Cocktail (Lablead, catalog number: R100).

Techniques: