human mouse ebi3  (Santa Cruz Biotechnology)

Journal: Scientific Reports

Article Title: Interleukin-35 administration counteracts established murine type 1 diabetes – possible involvement of regulatory T cells

doi: 10.1038/srep12633

Figure Lengend Snippet: ( A ) Concentrations of IL-10 and ( B ) IL-35 in supernatants of stimulated CD4 + CD25 + Treg cells sorted from thymic glands, PDLNS and spleen of vehicle or MLDSTZ treated mice. ( C ) Representative histograms showing the expression of IL-12p35 (left panels) or Ebi3 (right panels) in CD4 + CD25 + Foxp3 + Treg or CD4 + CD25 − Th cells in thymocytes, PDLNs cells and splenocytes of vehicle or MLDSTZ injected mice on indicated days. Results are expressed as means ± SEM, from two experiments (n = 3 mice/group/experiment). Unpaired t-tests were performed for comparisons between vehicle and MLDSTZ treated groups on corresponding days. *, ** and *** denote p < 0.05, p < 0.01, and p < 0.001, respectively.

Article Snippet: Ebi3 staining was made as follow: (1) The antigen retrieval was done by using Diva Decloaker buffer (BIOCARE MEDICAL) or Tris Buffered Saline buffer, pH 9.0. (2) Tissue sections were treated with 10% hydrogen peroxide to remove endogenous peroxidase. (3) Swine serum (1:20) was used to block the tissue sections (30 minutes). (4) Tissue sections were incubated overnight with anti-human/mouse Ebi3 (20 μg/ml; Novus Biologicals, Littleton, Colorado, USA) in a humidified chamber at 4 °C. (5) The next day, HRP polyclonal swine anti-rabbit antibody (1:200; Dako AB, Stockholm, Sweden) was applied for 60 minutes then visualized with DAB substrate (Sigma–Aldrich Sweden AB). (6) The sections were counterstained with Mayer’s hematoxylin (Histolab, Gothenburg, Sweden).

Techniques: Expressing, Injection

Journal: Scientific Reports

Article Title: Interleukin-35 administration counteracts established murine type 1 diabetes – possible involvement of regulatory T cells

doi: 10.1038/srep12633

Figure Lengend Snippet: Flow cytometry histograms showing the expression of Ebi3 in ( A ) CD4 + CD25 − T cells or ( B ) CD4 + CD25 + Foxp3 + Treg cells of thymus, PDLNs cells and spleen of MLDSTZ + PBS or MLDSTZ + IL-35 treated mice on day 14. ( C ) The IL-10 concentration was determined in the serum of different groups of MLDSTZ + PBS and MLDSTZ + IL-35 treated mice by using ELSA assay. ( D ) Histograms showing the expression of CD39 in CD4 + CD25 + Foxp3 + Treg cells of thymus, PDLNs cells and spleen of MLDSTZ + PBS or MLDSTZ + IL-35 treated mice on day 14. Results are expressed as means ± SEM (n = 6), from two experiments (n = 3 mice/group/experiment). One-way ANOVA followed by Shapiro-Wilk tests ( C ) were performed for comparisons between IL-35 and PBS treated mice, ** denotes p < 0.01.

Article Snippet: Ebi3 staining was made as follow: (1) The antigen retrieval was done by using Diva Decloaker buffer (BIOCARE MEDICAL) or Tris Buffered Saline buffer, pH 9.0. (2) Tissue sections were treated with 10% hydrogen peroxide to remove endogenous peroxidase. (3) Swine serum (1:20) was used to block the tissue sections (30 minutes). (4) Tissue sections were incubated overnight with anti-human/mouse Ebi3 (20 μg/ml; Novus Biologicals, Littleton, Colorado, USA) in a humidified chamber at 4 °C. (5) The next day, HRP polyclonal swine anti-rabbit antibody (1:200; Dako AB, Stockholm, Sweden) was applied for 60 minutes then visualized with DAB substrate (Sigma–Aldrich Sweden AB). (6) The sections were counterstained with Mayer’s hematoxylin (Histolab, Gothenburg, Sweden).

Techniques: Flow Cytometry, Expressing, Concentration Assay