APC anti-mouse CD194 (CCR4) 131211 from BioLegend

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Nature communications
June 10 2021

Fig 1: CCL2 binding affects the in vivo biodistribution of BC exosomes and facilitates BC metastasis.a, b DiD-labeled EO771 exosomes previously incubated (BC exo + CCL2, n = 6 animals) or not (BC exo, n = 8 animals) with CCL2 were injected into C57Bl/6 WT mice, and biodistribution assessed 24 h after injection. a Representative ex vivo images of DiD fluorescence in various organs from WT mice (clockwise from top-left: liver, spleen, kidney, lung, heart and bone marrow). b Quantification of DiD fluorescence in the same organs as shown in a (BC exo; black squares) (BC exo + CCL2; black triangles). c Frequency of CCR2+ population within lung (n = 12 animals), liver (n = 13 animals), bone marrow (n = 12 animals) and spleen (n = 13 animals) CD45.2+ cells as assessed by flow cytometry. d Frequency of CCR4+ and/or CCR2+ populations within CD45.2+ cells in the lung. e, f C57Bl/6 WT mice were injected with DiD-labeled EO771 exosomes previously incubated (BC exo + CCL2, n = 6 animals; black squares,) or not (BC exo, n = 9 animals; black triangles) with CCL2. e Frequency of DiD+ population within CD45.2+ cells in the lung. CCR2+ and CCR2- subpopulations were gated and analyzed separately. f Frequency of DiD+ population within distinct CD45.2+ immune cell subsets. CCR2+ and CCR2- subpopulations were gated and analyzed separately. g–i DiD-labeled EO771 exosomes previously incubated (BC exo + CCL2, n = 6 and n = 5 animals in the WT and CCR2-/- groups, respectively; black triangles) or not (BC exo, n = 6 and n = 9 animals in the WT and CCR2-/- groups, respectively; black squares) with CCL2 were injected into C57Bl/6 WT and CCR2-/- mice, and biodistribution assessed 24 h after injection. g Representative ex vivo images of DiD fluorescence in various organs from WT and CCR2-/- mice. h Quantification of DiD fluorescence in the lung as shown in g. i Frequency of DiD+ population within CD45.2+ cells in the lung of WT and CCR2-/- mice. j, k Mice pre-conditioned with cell culture-derived EO771 exosomes previously incubated (BC exo + CCL2, n = 7 and n = 5 animals in the WT and CCR2-/- groups, respectively; black triangles) or not (BC exo, n = 7 and n = 5 animals in the WT and CCR2-/- groups, respectively; black squares) with CCL2 received a single i.v. injection of EO771 cells, and metastatic burden in the lung was assessed. j Representative lung sections of each group (H&E staining). k Number of metastatic foci per tissue section (each data point represents a different mouse; one section was examined per mouse; all five lobes were analyzed and combined to determine total number of metastases per section). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 as analyzed by two-tailed Mann–Whitney U-test.
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Clinical and translational medicine
June 01 2022

Fig 2: Neutralisation of TGF-ß1 decreases lymphedema and inflammation. (A) Left panel: Representative cross-sectional H&E stain of a mouse tail treated with isotype control or TGF-ß1 neutralising antibody. Brackets show subcutaneous fibroadipose tissues; note decreased subcutaneous thickness in TGF-ß1 neutralising antibody-treated mice. Right panel: Quantification of skin thickness in control or TGF-ß1 neutralising antibody-treated mice. Each circle represents an average of three HPF views per animal (N = 4). (B) Left panel: Representative IF localisation of TGF-ß1 (top) and pSmad3 (bottom) in isotype and TGF-ß1 neutralising antibody-treated mice. Right panel: Quantification of TGF-ß1 area and Smad3+ cells. Each circle represents an average of three HPF views per animal (N = 5). (C) mRNA expression of TGF-ß1 isoforms and downstream signaling pathways. Relative change to isotype control-treated mice is shown. Each circle represents an individual animal (N = 5). Genes shown in the green- and red-shaded zones represent upregulated (green) and downregulated (red) molecules. *p < .05, **p < .01. (D) mRNA expression of inflammatory mediators. Relative change to isotype control-treated mice is shown. Each circle represents an individual animal (N = 5). Genes shown in the green- and red-shaded zones represent upregulated (green) and downregulated (red) molecules. *p < .05, **p < .01. (E) Quantification of flow cytometry for leukocytes (CD45+), CD45+/CD4+ cells, Th1 (CD45+/CD4+/CXCR3+/CCR5+) cells, Th2 (CD45+/CD4+/CCR4+/CCR8+) cells, macrophages (CD45+/CD11b+/F480+) and neutrophils (CD45+/CD11b+/Ly-6G+). Each circle represents an average of two flow cytometry runs for each animal (N = 5). (F) Quantification of CD4 T cells (CD45+/CD4+), macrophages (CD45+/CD11b+/F480+) and DCs (CD45+/CD11c+/CD86+) in draining lymph nodes and foot pad injection site after injection of LE lysate. Each circle represents an individual animal (N = 5). TGF-ß1, transforming growth factor-beta 1; H&E haematoxylin and eosin; HPF, high-power field; IF, immunofluorescence; DCs, dendritic cells; LE, lymphedema
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Clinical and translational medicine
June 01 2022

Fig 3: Topical PFD decreases the pathology of lymphedema. (A) Change in tail volume over time in mice treated with vehicle or PFD. Each circle is an average of duplicate measurements from each mouse (N = 9 animals per group). Results are presented as mean ± SEM (*p < .05, **p < .01). Statistical comparisons are between groups at the same time points. (B) Left panel: Representative cross-sectional H&E stain of vehicle and PFD-treated mouse tails. Brackets show subcutaneous fibroadipose tissue deposition. Right panel: Quantification of skin thickness in vehicle and PFD-treated mouse tails. Each circle represents an average of three HPF views from 5 animals. Bar = 500 µm. (C) Quantification of type I collagen deposition in vehicle and PFD-treated mouse tails. Each circle represents an average of three HPF views from 4 animals. (D) Quantification of TGF- ß1 protein in tissues collected from vehicle and PFD-treated mice. Each circle represents an average of duplicate ELISAs per animal (N = 9). (E) Quantification of the number of pSmad3+ cells in vehicle and PFD-treated mouse tails. Each circle represents an average of cell counts in three HPF views from 7 animals. (F) mRNA expression of TGF-ß1 isoforms and downstream signaling pathways. Relative change to vehicle control-treated mice is shown. Each circle represents an individual animal (N = 6). Genes shown in the green- and red-shaded zones represent upregulated (green) and downregulated (red) molecules. *p < .05, **p < .01, ***p < .001. (G) mRNA expression of fibrosis and ECM genes. Relative change to vehicle control-treated mice is shown. Each circle represents an individual animal (N = 6). Genes shown in the green- and red-shaded zones represent upregulated (green) and downregulated (red) molecules. *p < .05, **p < .01, ***p < .001. (H) mRNA expression of inflammatory cytokines. Relative change to vehicle control-treated mice is shown. Each circle represents an individual animal (N = 6). Genes shown in the green- and red-shaded zones represent upregulated (green) and downregulated (red) molecules. *p < .05, **p < .01, ***p < .001. (I) Quantification of flow cytometry for leukocytes (CD45+), macrophages (CD45+/CD11b+/F480+), CD45+/CD4+ cells, Th1 (CD45+/CD4+/CXCR3+/CCR5+) cells and Th2 (CD45+/CD4+/CCR4+/CCR8+) cells. Each circle represents an average of two flow cytometry runs per animal (N = 4). (J) Left panel: Representative IF localisation of CD4 (red) and LYVE-1 (green) in vehicle and PFD-treated mouse tails. Right panel: Quantification of perilymphatic CD4+ cells. Each circle represents an average of three HPF views from 9 animals. PFD, pirfenidone; SEM, standard error of the mean; H&E haematoxylin and eosin; HPF, high-power field; TGF-ß1, transforming growth factor-beta 1; ELISA, enzyme-linked immunosorbent assay; ECM, extracellular matrix molecules; IF, immunofluorescence

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APC anti-mouse CD194 (CCR4)

The APC anti-mouse CD194 (CCR4) Antibody from BioLegend is a Hamster Monoclonal antibody to CCR4. This antibody recognizes Mouse antigen. The APC anti-mouse CD194 (CCR4) Antibody has been validated for the following applications: Flow Cytometry.

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APC anti-mouse CD194 (CCR4) [2G12]

Nature communications

Clinical and translational medicine

Clinical and translational medicine

APC anti-mouse CD194 (CCR4) from BioLegend

Description

BioLegend

Citations (16)

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APC anti-mouse CD194 (CCR4) from BioLegend

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