Journal: JCI Insight

Article Title: Dendritic cell immunoreceptor drives atopic dermatitis by modulating oxidized CaMKII-involved mast cell activation

doi: 10.1172/jci.insight.152559

Figure Lengend Snippet: ( A and B ) Representative immunofluorescence images of dorsal skin sections and fluorescence analysis of DCIR staining in the skin tissues of patients with AD and controls ( n = 8). Scale bar: 100 μm. ( C ) Quantification analysis of DCIR + tryptase + cells in the lesion skin of patients with AD and controls. ( D ) Flow cytometry analysis of DCIR expression in human mast cell line cKit + FcεRI + LAD2 cells. ( E ) Scheme of experimental protocol for the direct bindings of human recombinant DCIR (hrDCIR) to BSA, CRE, and Man-BSA. ( F ) Direct binding of different doses of hrDCIR (0–5.0 μg/mL) to BSA, CRE, or Man-BSA ( n = 3). ( G ) Representative immunofluorescence images of FITC-CRE uptake by LAD2. Scale bar: 15 μm. ( H and I ) Flow cytometry analysis ( H ) and quantification ( I ) of FITC-CRE uptake at different doses (1–500 ng/mL) by LAD2 cells ( n = 3). ( J ) Inhibition of FITC-CRE uptake in LAD2 cells pretreated with DCIR neutralizing antibody (α-DCIR) or IgG isotype ( n = 4). Data represent mean ± SEM of 2 independent experiments. Data in B , C , and I were compared using a 2-tailed Student’s t test. Data in F and J were compared by 2-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The sections were then incubated with the primary antibodies against mouse Tryptase (AF1937, R&D system), DCIR (MAB2617, R&D), or mouse IgG1 overnight at 4°C.

Techniques: Immunofluorescence, Fluorescence, Staining, Flow Cytometry, Expressing, Recombinant, Binding Assay, Inhibition

Journal: JCI Insight

Article Title: Dendritic cell immunoreceptor drives atopic dermatitis by modulating oxidized CaMKII-involved mast cell activation

doi: 10.1172/jci.insight.152559

Figure Lengend Snippet: ( A ) Representative skin images and EASI scores of PBS- and CRE-treated WT and DCIR –/– mice. ( B ) Representative H&E staining and epidermal thickness (μm) of skin tissues of PBS- and CRE-treated WT and DCIR –/– mice. ( C ) Representative Toluidine blue staining and quantification of cells with positive staining for Toluidine blue of skin tissue sections of PBS- and CRE-treated WT and DCIR –/– mice. Scale bar: 100 μm. Arrows represent mast cells. ( D ) Serum levels of specific IgE and IgG1 to CRE. ( E ) Quantitative PCR analysis of IL-4, IL-13, IL-33, and TNF-α expression in the skin tissues of PBS- and CRE-treated WT and DCIR –/– mice. Each circle represents 1 mouse. n = 8. Data represent mean ± SEM of 2 independent experiments. Data were compared by 2-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The sections were then incubated with the primary antibodies against mouse Tryptase (AF1937, R&D system), DCIR (MAB2617, R&D), or mouse IgG1 overnight at 4°C.

Techniques: Staining, Real-time Polymerase Chain Reaction, Expressing

Journal: JCI Insight

Article Title: Dendritic cell immunoreceptor drives atopic dermatitis by modulating oxidized CaMKII-involved mast cell activation

doi: 10.1172/jci.insight.152559

Figure Lengend Snippet: ( A ) Scheme of experimental protocol of i.v. transfer of DCIR + versus DCIR – mast cells into Kit W-sh/W-sh mice for the generation of AD mouse model. ( B ) Representative Toluidine blue staining and quantification of cells with positive staining for Toluidine blue of skin tissue sections of Kit W-sh/W-sh mice with DCIR + or DCIR – mast cells ( n = 9). ( C ) Representative immunofluorescence images of mast cells with (yellow) or without (blue) DCIR expression. ( D ) Representative skin images and EASI scores of Kit W-sh/W-sh mice with DCIR + or DCIR – mast cells ( n = 8). ( E ) Representative H&E staining and epidermal thickness (μm) of skin tissues of Kit W-sh/W-sh mice with DCIR + or DCIR – mast cells ( n = 8). Scale bar: 100 μm. Arrows represent mast cells. ( F ) Serum levels of specific IgE and IgG1 to CRE ( n = 8). ( G ) Quantitative PCR analysis of IL-4, IL-13, IL-33, and TNF-α expression in the skin tissues of Kit W-sh/W-sh mice with DCIR + or DCIR – mast cells. Each circle represents 1 mouse ( n = 8). Data represent mean ± SEM of 2 independent experiments. Data were compared by 2-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The sections were then incubated with the primary antibodies against mouse Tryptase (AF1937, R&D system), DCIR (MAB2617, R&D), or mouse IgG1 overnight at 4°C.

Techniques: Staining, Immunofluorescence, Expressing, Real-time Polymerase Chain Reaction

Journal: Nucleic Acids Research

Article Title: Extensive remodeling of DC function by rapid maturation-induced transcriptional silencing

doi: 10.1093/nar/gku674

Figure Lengend Snippet: Identification of promoters undergoing H4-deacetylation upon Mo-DC maturation. ( A ) Schematic representation of the ChIP-chip strategy used to identify promoters that are deacetylated in Mo-DCs after 1 h of LPS treatment (top). Representative results for CIITA , IL12B , ACTB , CD1C , CD36 and CLEC4A are provided: signal ratios between 1 h-LPS-treated and untreated Mo-DCs are represented on a log 2 scale (bottom left). The percentages of promoters displaying increased or decreased H4Ac are shown (bottom right). ( B ) Gene-ontology analysis of genes exhibiting LPS-induced H4-deacetylation at their promoters was done using David ( http://david.abcc.ncifcrf.gov/ ). ( C ) H4-deacetylation (top), nascent transcripts (middle) and pol-II occupancy (bottom) were quantified for the indicated genes in untreated and 1 h-LPS-treated Mo-DCs: results are expressed relative to untreated DCs; nt, not tested. Statistical significance was derived from three experiments: *, P < 0.05, **, P < 0.01, ***, P < 0.001. All measurements were performed in triplicate for each experiment.

Article Snippet: Protein extracts were fractionated by Sodium dodecylsulphate-polyacrylamide gel electrophoresis and western blotting was performed using the following antibodies: MARCH1 (Abcam), CLEC10A (Abnova), SOCS5 (GeneTex), CLEC4A (GeneTex) and tubulin (SIGMA-ALDRICH).

Techniques: ChIP-chip, Derivative Assay