cd206 (mannose receptor) rat anti-mouse antibody  (Serono)

Journal: Marine Drugs

Article Title: Karenia brevis Extract Induces Cellular Entry through Distinct Mechanisms in Phagocytic RAW 264.7 Macrophages versus Non-Phagocytic Vero Cells

doi: 10.3390/md22010004

Figure Lengend Snippet: Assessment of macrophage alternative activation states by cytokine secretion (( A ) n-3) IL-10, TNF α and IL-6 and cell surface expression ( B ) of CD206 (n-10), CD80 (n-3) and IL4Rα (n-3). RAW 264.7 macrophages were treated with algal extracts for 24 h and stained with fluorescent antibodies prior to analysis on a flow cytometer. * indicates a statistically significant difference from vehicle control (VC) with p < 0.05.

Article Snippet: Antibodies included: (rat) anti-mannose receptor CD206 conjugated to Alexa Fluor 488 (R&D Systems, Minneapolis, MN, USA), (hampster) anti-mouse CD80 conjugated to PE-CF594 (BD Biosciences, San Jose, CA, USA), and (rat) anti-mouse IL4Rα conjugated to BV421 (BD Biosciences, San Jose, CA, USA).

Techniques: Activation Assay, Expressing, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Protease-Activated Receptor (PAR)2, but Not PAR1, Is Involved in Collateral Formation and Anti-Inflammatory Monocyte Polarization in a Mouse Hind Limb Ischemia Model

doi: 10.1371/journal.pone.0061923

Figure Lengend Snippet: (A) Stainings of CD206-positive macrophages (green) and SMA-positive vessels (red) in non-ischemic (control) and ischemic (ligated) hind limbs of WT, PAR1-/- and PAR2-/- mice are shown. Nuclei were visualized with DAPI (blue). Arrows indicate single macrophages in the non-ischemic adductor. Quantification of the average number of repair-associated macrophages per vessel is indicated on the right. (B) Correlation between the number of CD206-positive macrophages in the ischemic tissues and the expression of CD11b and (C) CD115 on monocytes. ** p<0.01, *** p<0.001.

Article Snippet: Specimens were blocked with PBS containing 2% fetal calf serum (FCS, PAA Laboratories, Cölbe, Germany) and 3% BSA followed by dual staining of arterial vessels and repair-associated macrophages using a Cy3-conjugated SMA antibody (clone 1A4, 1∶1000, Sigma-Aldrich) and a FITC-labelled mannose receptor CD206 antibody (clone MR5D3, 1∶50, AbD Serotec, Oxford, UK), respectively.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Estrogen Represses Hepatocellular Carcinoma (HCC) Growth via Inhibiting Alternative Activation of Tumor-associated Macrophages (TAMs) *

doi: 10.1074/jbc.M112.348763

Figure Lengend Snippet: Estrogen inhibits macrophage alternative activation in orthotopic mouse liver tumor models. A, tumors developed in the left hepatic lobe of BALB/c mice 6 weeks after intrahepatic inoculation of 106 Heps cells in each group. a–c, females. a, female control; b, females that underwent ovariotomy; c, E2 (50 μg/kg) administration after ovariotomy. d–f, males. d, male control; e, males that underwent castration; f, E2 (50 μg/kg) administration after castration (n = 8–9). In the OVX female mice, intrahepatic metastasis of liver cancer existed. B, average weights of tumors from each group. Tumors were stripped and weighed. The data are presented as mean ± S.E. (n = 8–9). In OVX mice, the liver tumor size was larger than in the intact mice. After administration of E2, tumor size for OVX mice was small compared with growth in OVX mice without E2 treatment. In male mice, we also found that estrogen administration after castration can result in relatively smaller liver tumors than in the castrated group. C, the macrophage cell numbers in tumor tissue were enumerated directly. Columns show the mean of three different experiments. Error bars, S.E. *, p < 0.05; **, p < 0.01. D, the ratio of CD206 and CD68 was used to demonstrate the relative cell numbers of CD206+ macrophage. E2 supplement decreased the CD206+ macrophage proportion compared with the surgical groups.

Article Snippet: CD206 (mouse macrophage mannose receptor) antibody was purchased from AbD serotec (Oxford, UK).

Techniques: Activation Assay, Control

Journal: The Journal of Biological Chemistry

Article Title: Estrogen Represses Hepatocellular Carcinoma (HCC) Growth via Inhibiting Alternative Activation of Tumor-associated Macrophages (TAMs) *

doi: 10.1074/jbc.M112.348763

Figure Lengend Snippet: Estrogen inhibits alternative activation of tumor-associated macrophage in vitro. A, E2 inhibits IL-4-induced arginase activity of ANA-1 cells. Values are expressed as means ± S.E. (n = 3 from three separate experiments; **, p < 0.01). B, E2 inhibits cocultured macrophage arginase activity. ANA-1 cells were pretreated with E2 for 2 days before coculture with Hepa1-6 cells. C, E2 inhibits cocultured macrophage mannose receptor expression in a dose-dependent manner. ANA-1 cells were pretreated with E2 for 2 days before coculture with Hepa1-6 cells. Anti-mouse CD206 (PE-conjugated; red) was used to label alternatively activated macrophage, and the percentage was calculated by flow cytometry analysis. Values are expressed as means ± S.E. (n = 3 from three separate experiments; *, p < 0.05). D–F, inflammatory cytokine protein secreted into culture medium. Values were expressed as means ± S.E. (n = 3 from three separate experiments; *, p < 0.05; **, p < 0.01; ***, p < 0.001). D, IL-10; E, IL-12p70; F, ratio of IL-10 to IL-12. G, E2 treatment could significantly inhibit IL-4-induced CD206 expression, as measured by flow cytometry analysis. H, Hepa1-6 cancer cell mobility is inhibited by E2-treated alternative macrophage. Mobility assays were carried out in 24-well Transwell units (cells per treatment condition in triplicate). Hepa1-6 cells were cocultured with no cells (a), control ANA-1 (b), alternatively activated ANA-1 (c), and E2-pretreated alternatively activated ANA-1 (d). After a 6-h incubation period, the moved cells that had passed through the membrane were stained and photographed (magnification, ×100). Error bars, S.E.; * and **, statistically significant p values <0.05 and <0.01, respectively.

Article Snippet: CD206 (mouse macrophage mannose receptor) antibody was purchased from AbD serotec (Oxford, UK).

Techniques: Activation Assay, In Vitro, Activity Assay, Expressing, Flow Cytometry, Control, Incubation, Membrane, Staining

Journal: The Journal of Biological Chemistry

Article Title: Estrogen Represses Hepatocellular Carcinoma (HCC) Growth via Inhibiting Alternative Activation of Tumor-associated Macrophages (TAMs) *

doi: 10.1074/jbc.M112.348763

Figure Lengend Snippet: Estrogen inhibits the downstream genes of the Jak1-Stat6 pathway. Total mRNA was extracted from ANA-1 cells treated with IL-4 and/or E2, and downstream genes Arg1 (A), Fizz1 (B), and CD206 (C) were analyzed by quantitative PCR. Error bars, S.E.

Article Snippet: CD206 (mouse macrophage mannose receptor) antibody was purchased from AbD serotec (Oxford, UK).

Techniques: Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Estrogen Represses Hepatocellular Carcinoma (HCC) Growth via Inhibiting Alternative Activation of Tumor-associated Macrophages (TAMs) *

doi: 10.1074/jbc.M112.348763

Figure Lengend Snippet: Estrogen exerts its inhibitory effects via ERβ but not ERα. The effects of estrogen receptor agonists on macrophage arginase activity were analyzed. Cells were precultured with E2 (nonspecific ER agonist), PPT (ERα-specific agonist) and DPN (ERβ-specific agonist) at the indicated concentration for 48 h and stimulated with IL-4 for another 24 h (A) or cocultured with Hepa1-6 (B). Values were expressed as means ± S.E. (error bars) (n = 6 from three separate experiments; *, p < 0.05 versus IL-4 treated group). C, Western blot analysis of total extracts from macrophages that were exposed to E2 (10 nm), ERα-specific agonist PPT (10 nm), and ERβ-specific agonist DPN (100 nm) for 24 h. Socs1 level was analyzed by immunoblotting after treatment with E2, DPN, and PPT. D, extracellular surface receptor CD206 of ANA-1 macrophage was detected by flow cytometry. Macrophages were pretreated with Estrogen receptor antagonist (ICI182,780) or siRNA to knock down ERβ (siESR2). The treatment in each group was independent from the others. siESR2 could abolish the inhibitory effects of estrogen.

Article Snippet: CD206 (mouse macrophage mannose receptor) antibody was purchased from AbD serotec (Oxford, UK).

Techniques: Activity Assay, Concentration Assay, Western Blot, Flow Cytometry, Knockdown