Journal: Blood

Article Title: Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages.

doi: 10.1182/blood-2009-07-231449

Figure Lengend Snippet: Figure 1. Galectin-5 is present on the surface of rat red cells. (A) Freshly isolated reticulocytes or erythrocytes were adsorbed on glass coverslips and processed for immunofluorescence as described in “Fluorescence-activated cell-sorting analysis of exosomes and red cells, fluorescence microscopy of red cells.” Transmission images of red cells (left) and corresponding fluorescence imaging (right) were recorded on cells by the use of purified rabbit anti–galectin-5 antibody followed by incubation withAlexa 488 anti–rabbit antibody. (B) Young reticulocytes and erythrocytes were analyzed by flow cytometry by the use of antibodies raised against Gal-2 (dashed line), Gal-4 (dotted line), and Gal-5 (solid line), already tested for their specificity (top), or the produced anti–galectin-5 serum (solid line) and the preimmune serum (bottom, dashed line). Tinted patterns indicate cell labeling obtained in the absence of primary antibodies. (C) Lymphocytes isolated from rat blood, as described in “Cells,” were analyzed by flow cytometry for Gal-5 (left, solid line), CD47 (middle, solid line) and Syto 16 green (right, solid line). Tinted patterns indicate cell labeling in the absence of primary antibodies. (D) Ghost and raft extracts isolated from reticulocytes or mature erythrocytes, as described in “Red cell subcellular fractionation,” were processed by SDS-PAGE and analyzed by Western blot for the indicated proteins. The molecular mass (kDa) standards are indicated on the left.

Article Snippet: Mouse anti–rat CD47 was from Serotec Limited.

Techniques: Isolation, Fluorescence, FACS, Microscopy, Transmission Assay, Imaging, Incubation, Cytometry, Produced, Labeling, Fractionation, SDS Page, Western Blot

Journal: Journal of neurochemistry

Article Title: Neuroinflammatory and behavioural changes in the Atp7B mutant mouse model of Wilson's disease.

doi: 10.1111/j.1471-4159.2011.07278.x

Figure Lengend Snippet: Fig. 2 Copper accumulation in liver and brain of toxic milk mice. (a, b) Copper content (a) and fold change (b) in copper in liver and brain areas of toxic milk mice in comparison to aged-matched controls. The number of livers was three per group and five brain tissues of different animals were pooled. Toxic milk mice and controls were matched for age and gender. Note differential change in different brain areas. Tx, toxic. Fig. 1 Livers of toxic milk mice show distinct histopathological chan- ges and inflammatory infiltrates. (a, b) Macroscopic pathology in livers (a) and HE stained liver sections (b) of toxic milk mice compared to age-matched controls. (c) Immunofluorescence or CD11b in liver sections of toxic milk mice and age-matched controls, demonstrating infiltrating lymphocytes. Scale bars in (b) and (c) are 100 lm. (d) Fold change in cytokine mRNA production in livers of toxic milk mice in comparison to controls. (e) Cytokine levels in the circulation of toxic milk mice compared to controls. Number of animals was five per group (three females, two males). Bars represent means + SEM. Asterisks indicate significant differences (*p < 0.05, ***p < 0.01). Tx, toxic.

Article Snippet: The following primary antibodies were used with respective concentrations: rabbit polyclonal anti-glial fibrillary acetic protein (GFAP) (1 : 1000, Dako, Glostrup, Denmark), rat monoclonal anti-mouse CD11b (1 : 200, Serotec, Oxford, UK) and mouse monoclonal against neuronal nuclei (NeuN) (1 : 500, Millipore, Temecula, CA, USA).

Techniques: Comparison, Staining

Journal: Journal of neurochemistry

Article Title: Neuroinflammatory and behavioural changes in the Atp7B mutant mouse model of Wilson's disease.

doi: 10.1111/j.1471-4159.2011.07278.x

Figure Lengend Snippet: Fig. 4 Inflammation in brains of toxic milk mice. (a–d) Astroglial (a) and microglial (c) reactivity in different brain regions of toxic milk and control mice as assessed by im- munohistochemistry for GFAP and CD11b, respectively, and quantification thereof (b, d). Scale bars in (a) are 100 lm, in (b) upper panels 200 lm and lower panels 500 lm. (e, f) Fold difference in cytokine, chemokine and inflammatory enzyme mRNAs in striatum (e) and hippocampus (f) of toxic milk and control mice. Number of animals was five per group (three females and two males per group). Significant dif- ferences are indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Str, striatum; Cc, corpus callosum; Tx, toxic.

Article Snippet: The following primary antibodies were used with respective concentrations: rabbit polyclonal anti-glial fibrillary acetic protein (GFAP) (1 : 1000, Dako, Glostrup, Denmark), rat monoclonal anti-mouse CD11b (1 : 200, Serotec, Oxford, UK) and mouse monoclonal against neuronal nuclei (NeuN) (1 : 500, Millipore, Temecula, CA, USA).

Techniques: Control

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

Article Title: Involvement of CD56brightCD11c+ cells in IL-18-mediated expansion of human γδ T cells.

doi: 10.4049/jimmunol.1001919

Figure Lengend Snippet: FIGURE 1. Effect of IL-18 on the growth of gd T cells and CD56brightCD11c+ cells. A, Effect of IL-18 (100 ng/ml) on expansion of gd T cells. PBMCs derived from a representative healthy adult individual were stimulated by ZOL (1 mM)/IL-2 (10 ng/ml) or 2M3BPP (100 nM–1 mM)/IL-2 (solid bar); ZOL/IL-2/IL-18 or 2M3BPP/IL-2/IL-18 (open bar); or ZOL/IL- 2/anti–IL-18Ra mAb (2 mg/ml; gray bar) or 2M3BPP/IL-2/anti–IL-18Ra mAb. The total number of gd T cells was quantified by trypan blue dye exclusion and flow cytometry. **p , 0.001. B, Massive cell aggregation induced by ZOL/IL-2/IL-18. Cell clusters were photo- graphed using a Nikon Digital Sight TE300- HM-2 (310) microscope (Nikon, Tokyo, Ja- pan) after 5 d of culture and analyzed by Lu- mina Vision Software (Mitani, Tokyo, Japan). C, Proportion of gd TCR-bearing cells in cul- ture on days 0 and 14. PBMCs were incubated in the presence of ZOL/IL-2/IL-18 for 14 d and analyzed for expression of CD3 and gd TCR. D, Phenotypic analysis of PBMCs in culture on days 0 and 14. PBMCs incubated with ZOL/ IL-2 or ZOL/IL-2/IL-18 were analyzed for expression of CD11c, CD3, and CD56 on days 0 and 14. CD56intCD11c2, CD56intCD11c+, CD56brightCD11c2, and CD56brightCD11c+ cells were gated, respectively. E, Time course of the expansion of gd T cells and CD56bright

Article Snippet: Depletion of CD3 +, CD56+, and CD14+ cells from PBMCs was conducted using LD columns and microbeads conjugated with mAbs to CD3, CD56, and CD14 (Miltenyi Biotec, Auburn, CA), respectively. gd T cells and CD14+ cells were purified by positive selection using MS columns (anti-TCR gd MicroBeads kit; Miltenyi Biotec).

Techniques: Derivative Assay, Cytometry, Microscopy, Software, Incubation, Expressing

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

Article Title: Involvement of CD56brightCD11c+ cells in IL-18-mediated expansion of human γδ T cells.

doi: 10.4049/jimmunol.1001919

Figure Lengend Snippet: FIGURE 2. Effect of depletion of CD56+ or CD14+ cells on the expansion of gd T cells. Whole PBMCs and PMBCs depleted of CD56+ or CD14+

Article Snippet: Depletion of CD3 +, CD56+, and CD14+ cells from PBMCs was conducted using LD columns and microbeads conjugated with mAbs to CD3, CD56, and CD14 (Miltenyi Biotec, Auburn, CA), respectively. gd T cells and CD14+ cells were purified by positive selection using MS columns (anti-TCR gd MicroBeads kit; Miltenyi Biotec).

Techniques:

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

Article Title: Involvement of CD56brightCD11c+ cells in IL-18-mediated expansion of human γδ T cells.

doi: 10.4049/jimmunol.1001919

Figure Lengend Snippet: FIGURE 3. Expansion of CD56brightCD11+ cells by IL-2/IL-18 in CD3-depleted PBMCs. A, Expan- sion of CD56brightCD11c+ cells in CD3-depleted PBMCs. CD3-depleted PBMCs were incubated in the presence of ZOL/IL-2 or ZOL/IL-2/IL-18 and then analyzed for expression of CD3, CD11c, and CD56. B, Proliferation of PBMCs depleted of CD3+ cells. CD3-depleted PBMCs were incubated in the presence of various cyto- kines in combinations, as indicated, and the number of living cells was counted at indicated time points. C, Expansion of CD56brightCD11c+ cells in CD3- depleted PBMCs. The total number of CD56bright

Article Snippet: Depletion of CD3 +, CD56+, and CD14+ cells from PBMCs was conducted using LD columns and microbeads conjugated with mAbs to CD3, CD56, and CD14 (Miltenyi Biotec, Auburn, CA), respectively. gd T cells and CD14+ cells were purified by positive selection using MS columns (anti-TCR gd MicroBeads kit; Miltenyi Biotec).

Techniques: Incubation, Expressing