goat anti mouse galectin 9 antibodies  (R&D Systems)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 binds to DR3. (a) Immunoprecipitation of recombinant human or mouse Galectin-9 with human DR3.Fc or 4-1BB.Fc. Control, human Galectin-3. (b) Immunoprecipitation of recombinant human or mouse Galectin-9 with mouse DR3.Fc. Observed molecular masses were h/m4-1BB.Fc or DR3.Fc – 50–58 kDa, m/hGalectin-9 – 36 kDa. Data in (a–b) are representative of at least three different experiments each. (c–f) Binding response of increasing concentrations (0.48–2000 nM) of hGalectin-9 (c, d) or mGalectin-9 (e, f) to immobilized hDR3.Fc (c, e) or mDR3.Fc (d, f), measured by SPR. Values represent average of three independent measurements. The response shown is reference-subtracted (unrelated Fc protein). (g–h) Human or mouse Galectin-9 or TL1A were coated onto ELISA plates and binding of WT or de-glycosylated DR3.Fc was detected using biotin anti-DR3 and streptavidin HRP. Competition was assessed with DR3.Fc first incubated with either TL1A or Galectin-9. Data are means ± s.e.m binding from triplicate cultures and representative of three different experiments.

Article Snippet: After SDS-PAGE, the proteins were visualized by western blotting with anti-mouse DR3 (R&D Systems) or anti-mouse Galectin-9 (BioLegend) followed by anti-Rat IgG light chain-specific-HRP (Jackson ImmunoResearch).

Techniques: Immunoprecipitation, Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Defective DR3 activity in T cells from Galectin-9−/− mice. (a) Naïve CD4+ T cells from WT or Galectin-9−/− mice were activated in vitro with anti-CD3 and anti-CD28 in the presence of control IgG and agonist TL1A.Ig. IL-2 and IFN-γ production were assessed after 48 h. (b) CD4+ T cells from WT and Galectin-9−/− mice were pre-activated with anti-CD3 and anti-CD28, and then after 48 h were restimulated with anti-CD3 in the presence of control IgG or agonist anti-DR3. IL-2 and IFN-γ production were assessed 48 h later. (c) CD4+ T cells from WT (solid line) and Galectin-9−/− (dotted line) mice were pre-activated as in (b) and stained for DR3 expression (pre-sort, left). Isotype control staining, shaded. Sorted cells were then restimulated with anti-CD3 in the presence of control IgG or agonist TL1A.Ig. IL-2 production was assessed at 24 h. All data are means ± s.e.m from triplicate cultures and representative of three different experiments. *p<0.05

Article Snippet: After SDS-PAGE, the proteins were visualized by western blotting with anti-mouse DR3 (R&D Systems) or anti-mouse Galectin-9 (BioLegend) followed by anti-Rat IgG light chain-specific-HRP (Jackson ImmunoResearch).

Techniques: Activity Assay, In Vitro, Staining, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Treg from Galectin-9−/− mice are refractory to stimulation through DR3. (a) CD4+Foxp3+ Treg cells from WT mice were stained for surface DR3 and intracellular Galectin-9. Shaded, isotype control. Dotted line, Treg from Galectin-9−/− mice. (b) Immunoprecipitation with TL1A.Ig or control Fc was performed on lysates from WT Treg cells. Immunoblotting was carried out with anti-DR3 (top) or anti-Galectin-9 (bottom). MW indicated. Data are representative of three experiments. (c) Treg cells from WT or Galectin-9−/− mice were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist TL1A.Ig. Proliferation was assessed at 72 h. Data are means ± s.e.m from triplicate cultures and representative of three experiments. *Significance TL1A.Ig WT vs. Galectin-9−/−. (d) WT (solid line) and Galectin-9−/− (dotted line) Treg cells were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist anti-DR3. After 48 h, cells were stained for intracellular IDO and IL-10. MFI indicated. Isotype controls, shaded. Data are representative of three different experiments. *p<0.05

Article Snippet: After SDS-PAGE, the proteins were visualized by western blotting with anti-mouse DR3 (R&D Systems) or anti-mouse Galectin-9 (BioLegend) followed by anti-Rat IgG light chain-specific-HRP (Jackson ImmunoResearch).

Techniques: Staining, Immunoprecipitation, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of EAE by anti-DR3. WT or Galectin-9−/− mice were immunized with MOG35-55 peptide and injected with either control IgG or agonist anti-DR3. (a) Percent Foxp3+CD4+ T cells in peripheral blood 5 days after the last injection of anti-DR3. (b) EAE clinical scores over time. (c) Frequencies of IL-17A+ CD4 T cells (left) and TNF+ CD4 T cells (middle and right) in draining lymph nodes on days 17 and 30, respectively. (d) Proportion of gated CD4+ cells expressing IL-10 and Foxp3 (left) and total numbers of Foxp3+IL-10+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. (e) Proportion of gated CD4+ cells expressing IL-17 or IFN-γ (left) and total numbers of IL-17+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. All data are either representative or means ± sem from five mice per group. Similar results in three different experiments. *p<0.05

Article Snippet: After SDS-PAGE, the proteins were visualized by western blotting with anti-mouse DR3 (R&D Systems) or anti-mouse Galectin-9 (BioLegend) followed by anti-Rat IgG light chain-specific-HRP (Jackson ImmunoResearch).

Techniques: Injection, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of allergic asthma by anti-DR3. WT and Galectin-9−/− mice were immunized with OVA to induce lung inflammation, and injected with IgG or agonist anti-DR3. (a) Representative H&E staining of lung sections (left), and mean inflammation score (right). (b) Eosinophil numbers and IL-5 expression in BAL. (c) Proportion of CD4+Foxp3+ T cells in BAL (left) and draining lymph nodes (right). All results are means ± s.e.m from five mice per group, and representative of three independent experiments. *p<0.05

Article Snippet: After SDS-PAGE, the proteins were visualized by western blotting with anti-mouse DR3 (R&D Systems) or anti-mouse Galectin-9 (BioLegend) followed by anti-Rat IgG light chain-specific-HRP (Jackson ImmunoResearch).

Techniques: Injection, Staining, Expressing

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Galectin-9 is bound to the surface of primary naive B cells. a Representative flow cytometry plot (left) and quantification (right) of geometric mean ± SEM of surface staining for galectin-9 in WT (black) and Gal9-KO (blue) B cells from nine independent experiments. b Representative DIC (left) and confocal microscopy images (right) mapped to an 8-bit fire color scale (ImageJ) of primary WT (top) and Gal9-KO B cells (bottom) stained for surface galectin-9. Quantification of number of galectin-9 puncta is shown on the right (each dot represents 1 cell, 20 cells measured per condition) with the mean ± SEM indicated by the red bar. Scale bar 2 μm. Data representative of three independent experiments. c Representative confocal microscopy images of cryosections of the inguinal lymph node of WT B cells stained for subcapsular sinus macrophages (CD169; blue), B cells (B220; magenta), and Gal9 (green). Scale bar 20 μm. Data representative of three independent experiments. Statistical significance was assessed by Mann-Whitney, **** p < 0.0001

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Flow Cytometry, Staining, Confocal Microscopy, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Galectin-9 regulates BCR microcluster formation and signaling. a Representative images of primary naive WT (top) and Gal9-KO B cells (bottom) fixed on bilayers containing anti-kappa as surrogate antigen (Ag) after 90 s of spreading and imaged by confocal microscopy. Brightfield (left) and confocal (right) visualizing antigen mapped to an 8-bit fire color scale (ImageJ). Scale bar 2 μm. Quantification of b area of spreading, c total antigen fluorescence intensity at the cell-bilayer contact, and d mean intensity of antigen for WT (black circles) and Gal9-KO (blue diamonds) cells (each dot represents 1 cell, 200 cells measured per condition), with the mean ± SEM indicated by the red bar, **** p < 0.0001, Mann-Whitney test. Data representative of at least three independent experiments. e – h Primary naive B cells from WT and Gal9-KO mice were settled onto anti-IgM-coated plates for the indicated time. Cells were lysed and subjected to SDS-PAGE followed by immunoblotting with e anti-phosphotyrosine and anti-ERK1/2, f anti-phospho-CD19, g anti-phospho-Akt, and h anti-phospho ERK1/2 (pERK) and anti-β tubulin. Data representative of at least three independent experiments. f Quantification of the fold increase in pCD19, pAkt, and pERK, with the mean ± SEM indicated by bar. Data were analyzed by two-way ANOVA, followed by Sidak’s multiple comparisons test; * p < 0.05, ** p < 0.01

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Confocal Microscopy, Fluorescence, MANN-WHITNEY, SDS Page, Western Blot

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Treatment with exogenous galectin-9 suppresses BCR signaling. a Representative flow cytometric histograms of WT (left) and Gal9-KO (middle) B cells either untreated or treated with various concentrations of recombinant galectin-9 (rGal9; 0.1, 0.2, 0.5, and 1 μM) followed by surface staining for galectin-9 and analyzed using flow cytometry. Overlay of endogenous galectin-9 surface expression in WT cells, and Gal9-KO cells treated with 0.1 μM rGal9 (right). b Naive B cells from WT and Gal9-KO mice treated with 0.1 μM rGal9 were settled onto anti-IgM-coated plates for the indicated time. Cells were lysed and subjected to SDS-PAGE followed by immunoblotting with anti-phospho ERK1/2 and anti-β tubulin (left panel). Quantification of the fold change in pERK over time, averaged over two independent experiments with the mean ± SEM indicated by the bar (right panel). c – f Naive B cells from WT mice were treated with 1 μM rGal9 and settled onto anti-IgM-coated plates for the indicated time. Cells were lysed and subjected to SDS-PAGE followed by immunoblotting with c anti-phosphotyrosine and ERK1/2, d anti-phospho-CD19, e anti-phospho-Akt, and f anti-phospho ERK1/2 and anti-β tubulin. Quantification of the fold change in pCD19, pAkt, and pERK over time, averaged over three independent experiments, with the mean ± SEM indicated by the bar is shown in the right panel. Statistical significance measure by two-way ANOVA followed by Sidak’s multiple comparisons test; **** p < 0.0001, ** p < 0.01, * p < 0.05

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Recombinant, Staining, Flow Cytometry, Expressing, SDS Page, Western Blot

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Galectin-9 alters IgM-BCR nanoclusters. a TIRFM image of surface IgM and fluorescently labeled rGal9 before bleaching for image acquisition (two left panels respectively). dSTORM images reconstructed from single-molecule localization processed by Thunderstorm software mapped to a fire color scale as indicated; the magnified region (3 × 3 µm) from ROI (white box) is shown as 2D image (middle) and 3D surface plot (right) in the order of WT (top), Gal9-KO (middle), and Gal9-KO + 1 µM rGal9 (bottom). Scale bar represents 2 µm. b Quantification of the distribution of IgM by H function and c Hopkins index of localizations inside ROIs. d – g Reconstructed images were analyzed by a model-based Bayesian approach to identify nanoclusters and their physical properties. d Number of clusters (one point per ROI). e Cluster radii (one point per cluster). f Number of molecules (one point per cluster). g Percentage of localization in clusters (one point per ROI). Each category contains at least 15 ROIs from three independent experiments (at least four cells per experiment). Statistical analysis was performed using Kruskal-Wallis test with Dunn’s multiple comparison test ( d , e , f ) and one-way ANOVA with Tukey’s multiple comparison test ( b , g ). Red bars indicate mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Labeling, Software, Comparison

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Galectin-9 immobilizes IgM-BCR and attenuates BCR microclustering. a Diffusion coefficients and b frequency distribution histogram of single-particle tracking of IgM in WT (black circle) or Gal9-KO (blue diamond) primary B cells with the median indicated in red. Five hundred representative diffusion coefficients from a total of at least 1500 tracks from three independent experiments. c Representative TIRF image of fluorescently labeled rGal9 on primary B cell (left) and mask (right) created to differentiate tracks inside Gal9 regions (lower left, red lines) and tracks outside Gal9 regions (lower right, yellow lines). d Diffusion coefficients and e frequency distribution inside Gal9 regions (black circle) and outside Gal9 regions (black triangle) with the median indicated in red. In all, 250 representative diffusion coefficients from a total of at least 900 tracks from three independent experiments. f Representative TIRF microscopy images of WT cell (top) and WT cells treated with 1 μM rGal9 (bottom) on artificial planar lipid bilayers containing anti-kappa mapped to an 8-bit fire color scale (ImageJ). g Quantification of the total antigen intensity at the cell-bilayer interface in WT (black circles) and WT treated with rGal9 (open circles), with the mean ± SEM indicated by the red bar. Scale bar represents 2 μm. Statistical significance assessed by Mann-Whitney; **** p < 0.0001, *** p < 0.001, ** p < 0.01

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Diffusion-based Assay, Single-particle Tracking, Labeling, Microscopy, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: The galectin-9 lattice increases the molecular density of IgM-BCR and co-receptors. a Representative confocal images of primary WT B cells treated with 1 µM rGal9 and immuostained for CD45 (cyan), IgM (magenta), and galectin-9 (Gal9; yellow). b Fluorescence intensity profile of CD45, IgM, and Gal9 along the cell membrane. c Representative example of masking output of algorithm to detect regions of high galectin-9 (Gal9 high ) and low galectin-9 (Gal9 low ). d Mean fluorescence intensity of CD45 (left) and IgM (right) in Gal9 high and Gal9 low regions. e Representative confocal images of WT B cells treated with 1 µM rGal9 and immunostained for CD22 (cyan), IgM (magenta), and Gal9 (yellow). f Fluorescence intensity profile of CD22, IgM, and Gal9 along the cell membrane. g Mean fluorescence intensity of CD22 (left) and IgM (right) in Gal9 high and Gal9 low regions. h Representative confocal images of WT B cells treated with 1 μM rGal9 and immunostained for CD19, IgM, and Gal9. i Fluorescence intensity profile of CD19, IgM, and Gal9 along the cell membrane. j Mean fluorescence intensity of CD19 (left) and IgM (right) in Gal9 low and Gal9 high regions. Data representative of at least three independent experiments. Each dot represents 1 cell, at least 30 cells measured per condition per experiment. Mean ± SEM indicated by the red bar. Statistical significance assessed by Mann-Whitney; **** p < 0.0001 *** p < 0.001, * p < 0.05. Scale bar 2 μm

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Fluorescence, Membrane, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: rGal9 induces coalescence of lipid raft domains containing CD22 and CD45. a Representative confocal images of primary WT B cells treated with 1 μM rGal9 and immunostained for CD45 (cyan) and galectin-9 (Gal9; yellow), and fluorescent cholera toxin (CT-B; gray) to label lipid rafts. b Fluorescence intensity profile of CD45, CT-B, and Gal9 along the cell membrane. c Representative example of masking output of algorithm to detect regions of high CT-B (lipid raft high; LR high ) and low CT-B (LR low ). d Mean fluorescence intensity of CD45 (left) and Gal9 (right) in LR low and LR high regions. e Representative confocal images of WT B cells treated with 1 μM rGal9 and immunostained for CD22 (cyan), Gal9 (yellow), and fluorescent CT-B (gray). f Fluorescence intensity profile of CD22, CT-B, and Gal9 along the cell membrane. g Mean fluorescence intensity of CD22 (left) and Gal9 (right) in LR low and LR high regions. Data are representative of at least three independent experiments. Each dot represents 1 cell, at least 30 cells measured per condition per experiment. Mean ± SEM indicated by the red bar. Statistical significance assessed by Mann-Whitney; **** p < 0.0001, *** p < 0.001. Scale bar 2 μm

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Fluorescence, Membrane, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Galectin-9 increases colocalization between CD22 and IgM in primary B cells. a Representative merged TIRF (top), dSTORM (middle), and dSTORM zoom (bottom) images showing surface CD45 (magenta) and IgM-BCR (green) on primary wild-type (WT) (left) and galectin-9 knockout (Gal9-KO) (right) B cells. dSTORM ROI (3 × 3 μm) is outlined in yellow (middle) and magnified in dSTORM zoom (bottom). b – e Quantification of at least 20 ROIs from WT and Gal9-KO B cells pooled from three independent experiments. b Hopkin’s index showing randomness of CD45 organization (one point per ROI). c H function derived from Riplely’s K showing degree of CD45 clustering. d Mean diameter of CD45 clusters (one point per ROI). e Mean area of CD45 clusters (one point per ROI). f , g Quantification of at least 15 ROIs from WT and Gal9-KO B cells pooled from three independent experiments. f Coordinate-based colocalization (CBC) histograms of the single-molecule distributions of colocalizations between CD45 and IgM. g Nearest-neighbor distance (NND) analysis of the data shown in f . Symbol represents the median NND of all paired single-molecule localizations from one ROI. h Representative merged TIRF and dSTORM images showing surface CD22 (magenta) and IgM-BCR (green) on primary WT (left) and Gal9-KO (right) B cells. i – l Quantification of at least 30 ROIs from WT and Gal9-KO B cells pooled from three independent experiments. i Mean Hopkin’s index showing randomness of CD22 organization (one point per ROI). j H function showing degree of CD22 clustering. k Mean diameter of CD22 clusters (one point per ROI). l Mean area of CD22 clusters (one point per ROI). m , n Quantification of at least 20 ROIs from WT and Gal9-KO B cells pooled from three independent experiments. m CBC histograms of the single-molecule distributions of colocalizations between CD22 and IgM. n NND analysis of the data shown in m . Colocalization between channels shown in white. Scale bars represent 2 and 1 μm (zoom). Mean ± SEM indicated by the red bar. Statistical significance assessed by Mann-Whitney, * p < 0.05

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Knock-Out, Derivative Assay, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-9 binds IgM-BCR to regulate B cell signaling

doi: 10.1038/s41467-018-05771-8

Figure Lengend Snippet: Schematic model of galectin-9 regulation of B-cell activation. a In resting primary naive WT B cells, galectin-9 facilitates interactions between BCRs and either the inhibitory proteins CD45 or CD22 through binding to N-linked glycans, providing a basal attenuation of B-cell signaling upon antigen stimulation. b BCR signaling is enhanced in Gal9-KO B cells due to loss of association of BCR with inhibitory co-receptors. c Treatment of WT B cells with rGal9 induces the association of IgM-BCR with CD45 and CD22 to suppress B-cell signaling

Article Snippet: Cells were incubated with 1 μg/mL goat anti-mouse Gal9 (R&D systems, Cat. No. AF3535) in FACS buffer for 1 h at 4 °C.

Techniques: Activation Assay, Binding Assay

Journal: PLoS ONE

Article Title: Upregulation of the Tim-3/Galectin-9 Pathway of T Cell Exhaustion in Chronic Hepatitis B Virus Infection

doi: 10.1371/journal.pone.0047648

Figure Lengend Snippet: PBMC derived from HLA A2+ patients with CHB were stained with a panel of HLA-A2/HBV multimers and then were stimulated overnight with a pool of HBV peptides of matched specificity to the multimers, followed by intracellular staining for IFN-γor TNF-α. (a) Representative histograms showing levels of Tim-3 (black line) or isotype binding (grey shading) on CD8 T cells binding HLA-A2/HBV peptide multimers or producing IFN-γ upon encounter with HBV peptides. (b) Compiled data from 10 patients with CHB. (c) Tim-3 expression on CD8 T cells binding HLA-A2/HBV peptide multimers or producing TNFa upon stimulation with HBV peptides. (d) FACS plots and (e) summary data showing the induction of caspases (FLICA) and 7AAD in CD8 and CD4 T cells with or without the addition of galectin-9. Active caspases, indicating apoptosis, were determined using a fluorescent-labelled inhibitor of polycaspases (FAM-VAD-FMK, FLICA), and death was identified by 7AAD stain. Early apoptotic events are indicated in the lower right quadrant (FLICA+7AAD−), late apoptotic events in the right upper quadrant (FLICA+7AAD+) and necrotic cells in the left upper quadrant (7AAD+FLICA-). ‘Total death’ was estimated by summing events in these 3 quadrants.

Article Snippet: Immunostaining of cryopreserved sections from HBV infected liver biopsies was performed using a combination of the following primary antibodies Galectin-9 goat polyclonal IgG (R & D Systems) and CD68 mouse monoclonal (KP1, Abcam).

Techniques: Derivative Assay, Staining, Binding Assay, Expressing

Journal: PLoS ONE

Article Title: Upregulation of the Tim-3/Galectin-9 Pathway of T Cell Exhaustion in Chronic Hepatitis B Virus Infection

doi: 10.1371/journal.pone.0047648

Figure Lengend Snippet: Immunohistochemistry of a section from a cryopreserved CHB liver biopsy stained with a polyclonal galectin-9 antibody (shown in brown) at 10X and 40X magnification. (b ) Immunofluorescence of a section from a cryopreserved CHB liver biopsy stained with DAPI (blue, left panel), anti-CD68 mAb (green, central panel) and galectin 9 polyclonal antibody (red, right panel). Double positive staining is indicated in yellow in the lower panel. (c) Kupffer cells were isolated from 4 liver explants and stained with CD14, CD68 and galectin-9 or its isotype. Representative histograms show galectin-9 staining on CD14+CD68+ fraction compared to isotype-matched control. (d) Galectin-9 levels were analysed in the serum of 10 healthy subjects and 42 CHB patients: 16 with ALT<50, 17 with ALT 50–100 and 9 with ALT >100 (mean and SEM shown).

Article Snippet: Immunostaining of cryopreserved sections from HBV infected liver biopsies was performed using a combination of the following primary antibodies Galectin-9 goat polyclonal IgG (R & D Systems) and CD68 mouse monoclonal (KP1, Abcam).

Techniques: Immunohistochemistry, Staining, Immunofluorescence, Isolation