recombinant ccl8 protein  (MedChemExpress)

Journal: bioRxiv

Article Title: Immune disease dialogue of chemokine-based cell communications as revealed by single-cell RNA sequencing meta-analysis

doi: 10.1101/2024.07.17.603936

Figure Lengend Snippet: a) Pan CD8 T cells migrated towards CXCL2 (at all dilutions for donor 1, and at 1 ng/ml and 10 ng/ml for donor 2), b) pan monocytes did not migrate towards RARRES2, c) pan monocytes migrated towards CCL8 (at 0.1 ng/ml, 1 ng/ml and 100 ng/ml for donor 1, and at 0.1 ng/ml and 100 ng/ml for donor 2).* p < 0.05. ** p < 0.01, *** p < 0.0001 compared to the medium control condition per donor.

Article Snippet: For the monocyte experiments, two positive controls were used; 5% (v/v) cobra venom activated human complement serum (CAS; Complement Technology Inc, cat. NC1769554), as well as CC motif chemokine ligand 8 (CCL8; R&D Systems, cat. 281-CP-010).

Techniques: Control

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Single-cell transcriptomic landscape of subcutaneous LLC tumors at early-stage following hypofractionated radiotherapy. A Overview of the experimental design for single-cell RNA sequencing. B Uniform Manifold Approximation (UMAP) plot showing the unsupervised clusters of 54883 single cells and annotated cell types. C The proportion of each cell types in LLC tumors treated with or without radiation. For B and C, each color represents the same cell type. D Feature plot and E Dot plot showing marker genes. F Heatmap showing serum chemokines level in LLC murine models 72 h post-treatment. Each row in the heatmap has been scaled. G The concentrations of CCL2, CCL7, CCL8, and CSF1 in serum measured by ELISA (n = 5/group). Data are presented as mean ± SD with student unpaired t test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant

Article Snippet: Recombinant CCL8 protein was purchased from MCE (Cat. #: HY-P7771).

Techniques: RNA Sequencing, Marker, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Identification of M2-like Ccl8 high Macrophages in the LLC-bearing murine model. A UMAP plot showing the annotation of macrophage populations in LLC tumors. B Feature plot and C Dotplot showing marker genes of macrophage populations. D Cell proportion of each cell type in NT and RT groups. E Heatmap displaying scores of M1, M2, angiogenesis, phagocytosis for each macrophage population. F Correlation analysis of CCL8 and CD163 expression level in TCGA-LUAD datasets (Spearman’s rho value = 0.542, p < 0.001). G The Gene Ontology (GO) enrichment analysis of each macrophage populations. H Heatmap displaying transcription factors activity of the Mac_Ccl8, Mac_Hmox1, Mono_Cxcl3, and Mono_Plac8 populations. I Development trajectory of macrophages populations predicted by Monocle2. J The cell density and K gene expression patterns along with the pseudotime,

Article Snippet: Recombinant CCL8 protein was purchased from MCE (Cat. #: HY-P7771).

Techniques: Marker, Expressing, Activity Assay, Gene Expression

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy promoted the crosstalk between the Mac_Ccl8 and lymphocytes. A UMAP plot showing the annotation of lymphocyte populations. B Feature plot and C Dot plot showing marker genes of lymphocyte populations. D The proportion of each lymphocyte population in two groups. E Violin plots displaying the expression level of immune checkpoint ligand genes in each lymphocyte populations of NT and RT groups. F Circle plots showing number of interactions in two groups inferred by CellChat. G The comparison of cellular communication probability from Mac_Ccl8 to T and NK cells in between two groups. H Chord plot displaying the upregulated signaling pathways in the RT group relative to the NT group. I Heatmap of the differential interaction strength of the GALECTIN signaling pathway between two groups

Article Snippet: Recombinant CCL8 protein was purchased from MCE (Cat. #: HY-P7771).

Techniques: Marker, Expressing, Comparison, Protein-Protein interactions

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy reprograms CCL8 high macrophages through the CCL signaling pathway. A Volcano plot showing differentially expressed genes of the Mac_Ccl8 between the NT and RT groups. Adjusted p value < 0.05, two-sided Wilcoxon test. B Violin plots comparing the expression of Ccl2, Ccl3, Ccl4, Ccl7, Ccl8, and Ccl12 in the NT and RT groups. Unpaired two-sided Wilcoxon test. C Representative examples of multiplex immunofluorescent labeling CD206 and CCL8. Green, CD206; Red, CCL8; Blue, DAPI. D Bar plots showing the GO enrichment analysis of upregulation and downregulation genes in the RT group. E Differences in IFN-Gamma and TNF pathways activity between two groups inferred by GSEA. F Cell–cell communication network between myeloid populations and LLC cells. G River plot displaying communication patterns of different cell types. H Chord plot (top) and heatmap (bottom) showing communication network of CCL signaling pathway in different cell types. I Weighted network analysis of differential interaction strength of signals in the Mac_Ccl8 population between two groups. J The comparison of cellular communication probability from Mac_Ccl8 to other myeloid populations between two groups. K Chord plot displaying the upregulated signaling pathways in the RT group relative to the NT group. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant

Article Snippet: Recombinant CCL8 protein was purchased from MCE (Cat. #: HY-P7771).

Techniques: Expressing, Multiplex Assay, Labeling, Activity Assay, Comparison, Protein-Protein interactions

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy promotes M2-like Ccl8 high macrophages infiltration and leads to poor prognosis. A Kaplan–Meier plots showing worse clinical prognosis in the LUAD and HNSC patients with the higher expression level of the Mac_Ccl8 signature. HR, hazard ratio. B Schematic diagram of the combination treatment with hypofractionated radiotherapy and the Bindarit. The intraperitoneally administration of Bindarit began from day 5 to day 11 post-tumor injection, and radiation treatment was initiated from day 7 to day 9 post-tumor injection. C Growth curves of tumors in LLC-bearing mice in the indicated treatment groups (n = 5 mice/group). Data are presented as mean ± SD with two-way ANOVA test. D Representative examples in the indicated treatment groups of multiplex immunofluorescent labeling F4/80, CD206 and CCL8. Yellow, F4/80, Green, CD206; Red, CCL8; Blue, DAPI. E Percentages of M1 and M2 macrophages in the specific treatment groups analyzed by flow cytometry (n = 3/group). F Representative flow cytometry panels showing M2 macrophages (top) and M1 macrophages (bottom). BI, Bindarit; RT, Radiation therapy; RT + BI, the combination therapy of the radiation and the Bindarit

Article Snippet: Recombinant CCL8 protein was purchased from MCE (Cat. #: HY-P7771).

Techniques: Expressing, Injection, Multiplex Assay, Labeling, Flow Cytometry

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: Primer sequences.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques: Sequencing

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: Antibody list.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques:

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: CCL8 was induced by lactate in macrophages. ( A , B ) KEGG pathway and volcano map and volcano map analysis of differential genes on lactate-treated macrophages for 24 h. ( C ) qRT-PCR in macrophages stimulated with CM3 or LA for 24 h. ( D ) ELISA detection of CCL8 in the serum of healthy and CRC donors. ( E , F ) Representative images and quantitative analysis of immunohistochemistry for CCL8 and CD68 in CRC tissues and adjacent nontumor tissues. All t -tests were two-tailed. Mean ± SEM. ** p < 0.01; *** p < 0.001. Scale bar: 100 μm.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Two Tailed Test

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: RNA sequencing analysis of M2-macrophage-related chemokines in macrophages treated with lactate for 24 h.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques: RNA Sequencing

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: ( A ) Western blot analysis of CCR5 in CRC tissue and adjacent nontumor tissue. ( B , C ) Representative images and quantitative analysis of the colony assay in HCT-116 and RKO cells treated with PBS, CCL8, or CCL8+Maraviroc for 15 days. ( D , E ) Representative images of the immunohistochemistry for skeleton proteins (F-actin and p-SMAD2) in RKO and HCT-116 cells treated with PBS, CCL8, or CCL8+Maraviroc for 24 h. ( F ) Western blot analysis of EMT-related proteins (PCNA, vimentin, N-cadherin) in RKO cells stimulated with CCL8, Maraviroc, or both for 24 h. ( G – K ) Representative images and quantitative analysis of subcutaneous and lung metastasis models of Balb/c-nude tumor-bearing mice, treated with PBS, CCL8, or CCL8 + Maraviroc. All t -tests were two-tailed. Mean ± SEM. * p < 0.05, ** p < 0.01; *** p < 0.001; ns, no significance. Original western blots are presented in . Scale bar: 100 μm.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques: Western Blot, Colony Assay, Immunohistochemistry, Two Tailed Test

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: ( A – C ) The alterations of the mTOR/4EBP1/70S6K signaling pathway in RKO cells, after treatment with CCL8, Maraviroc, or a series of combinations of CCL8 and Maraviroc for 24 h. ( D ) The alterations in the mTOR/4EBP1/70S6K signaling pathway in RKO cells before and after CCR5 knockdown, treated with or without CCL8 for 24 h. Original western blots are presented in .

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques: Knockdown, Western Blot

Journal: Cancers

Article Title: Lactate-Induced CCL8 in Tumor-Associated Macrophages Accelerates the Progression of Colorectal Cancer through the CCL8/CCR5/mTORC1 Axis

doi: 10.3390/cancers15245795

Figure Lengend Snippet: Schematic model showing the tumor-progression-promoting interaction between CRC cells and TAMs. Tumor-associated macrophages (TAMs) are more likely to adopt an M2-type polarization in response to the high lactate level in the tumor microenvironment. This leads to an increase in the secretion of CCL8 by polarized TAMs, which binds to the CCR5 receptor on tumor cells and promotes their proliferation and metastasis via the mTOR/70S6K/4EBP1 signaling pathway.

Article Snippet: Recombinant human CCL8 (rCCL8) protein, macrophage colony-stimulating factor (MCSF), and Maraviroc (selective antagonist of CCR5) were purchased from MCE.

Techniques:

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: M1 and M2 macrophages were co-cultured with BLM melanoma cells for transcriptomic (24 h) and secretomic (72 h) analyses. (A) Gene set enrichment analyses (GSEA) comparing 24 hours exposed vs un-exposed isolated cells, as indicated (n=3 donors). Normalized Enrichment Score (NES) values from the main upregulated hallmark_pathwaysfor all groups,are shown. GOMF and KEGG gene sets were additionally studied for cytokine/ chemokine inflammatory pathways. False Discovery Rates are represented (FDR q value <0.01). (B) Representative scheme highlighting the coincident most upregulated secreted chemokines (fold >4 to both unex-posed macrophages and melanoma cells). (C) Secretomic analysis (38 chemokines) of 72 hours co-cultured supernatants comparing M1+ BLM or M2+BLM with unexposed macrophages (x-axis) or BLM (y-axis) cells. Fold logarithmic average changes are shown (n=4 donors). (D) CCL8 and CCL15 secretion assessed at 72 h macrophages ±BLM conditioned media. Mean ±standard deviation (SD) values are shown (n= 5 do-nors). (E) CCL8 and CCL15 mRNA expression levels in mutually conditioned isolated cells. Mean ±SD values relative to TBP mRNA are shown (n= 4). Significant differences to the respective untreated controls, are shown (*p< 0.05, unpaired t-test).

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: Cell Culture, Isolation, Standard Deviation, Expressing

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: (A) Serum deprivation survival assays of BLM, A375 and Skmel-103 melanoma cell lines in response to increasing concentra-tions of CCL8 and CCL15 chemokines (ng/ml), and to FCS 1%, as positive control. Mean ±SD optical density (OD) values, are shown (n= 4). (B) Proliferation response of melanoma cell lines to 200 ng/ml exogenous CCL8 and CCL15, and to FCS 1% as positive control. Mean ±SD percentages of BrdU+ nuclei, are shown (n= 3). (C) BLM spheroids embedded in 3D-collagen and allowed to invade for 72 h in the presence of increasing concentrations of exogenous CCL8 and CCL15, as indicated (ng/ml). Functional chemotactic axes were identified by using neutralizing antibodies against CCR1, CCR3, CCR5 and their corresponding control isotypes (5 μg/ml). Mean ±SD fold-invasion values, are shown (n= 3-6). Images show representative collagen-embedded spheroids showing basal and CCL15 stimulated invasion. Scale bar, 150 μm. (D) Immunoblots showing CCR1, CCR3 and CCR5 expression in whole-lysates of three melanoma cell lines and isolated CD14+ monocytes. (E) Serum deprivation survival and 3D-collagen invasion assays of monocytes in response to 100 ng/ml exogenous chemokines, including FCS 1%, as positive control (n= 5 donors). Significant differences to the respective untreated controls, are shown (*p< 0.05, paired t-test).

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: Positive Control, Functional Assay, Control, Western Blot, Expressing, Isolation

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: TAM (A) and tumor cell (B) average MFI of CCL15, CCL8, CCR1, CCR3 and CCR5 in non-metastasizing and metastasizing primary tumors.Mann-Whitney statistical analysis was used to compare non-metastasizing vs metastasizing melanomas; p-values are shown.(C) FFPE human melanoma samples stained for CD68 (TAM marker, red) and CCL15, CCL8, CCR1, CCR3 or CCR5 (green). Scale bar, 50 μm.

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: MANN-WHITNEY, Staining, Marker

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: (A)Disease-free and overall survival 10-year Kaplan–Meier curves. Youden’s index was used to choose a cutoff point to classify the 67 primary melanomas as ‘low’ or ‘high’: TC CCL8 (MFI, 80 a.u.), TC CCL15 (98a.u.),TAM CCL15 (69 a.u.) and TAM CCR3 (78 a.u.). Log-rank p values, are shown. (B) Summarizing model, created with BioRender.com .

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: