Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: IL-4 enhances IL-12 p70 production by mouse splenic DCs. Mouse splenic DCs were cultured with an IL-12 stimulus (LPS, anti-CD40 mAb, poly I:C, CpG, or SAC [10 μg/ml], or a mix of all stimuli together) in the presence of GM-CSF and IFN-γ with or without IL-4, for 18 h. The supernatants were assayed for IL-12 p70 content by ELISA. The result is typical of more than five experiments of this type.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: Titration of the effect of IL-4 on the production of IL-12 by mouse splenic DCs. Mouse splenic DCs were cultured with a mix of stimuli as used in (but with SAC used at only 5 μg/ml), in the presence of IFN-γ and GM-CSF and titrated concentrations of IL-4, for 18 h. A neutralizing mAb to IL-4 (10 μg/ml) was included for one set of conditions as indicated (gray bar). The supernatants were analyzed for IL-12 p70 and p40 by ELISA (A and B) and Western blotting (C). The result is typical of three similar experiments with ELISA readouts and Western blotting.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Titration, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: The effects of combinations of IL-4, GM-CSF, and IFN-γ on the IL-12 production by mouse splenic DCs. Mouse splenic DCs were cultured with a mix of stimuli as used in in the presence or the absence of IL-4, IFN-γ, or GM-CSF, for 23 h. The supernatants were analyzed for IL-12 p70 and p40 by ELISA (A and B) and Western blotting (C). The result is typical of more than five similar experiments with ELISA readouts and Western blotting.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: Titration of the effect of IFN-γ on the production of IL-12 by mouse splenic DCs. Mouse splenic DCs were cultured with SAC (20 μg/ml) in the presence of titrated concentrations of IFN-γ, for 23 h. The supernatants were analyzed for IL-12 p70 and p40 by ELISA (A and B) and Western blotting (C). The error bars represent the variation within one typical experiment. The experiment has been repeated three times for the ELISA and two times for Western blots, with similar results.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Titration, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: Titration of the effect of GM-CSF on the production of IL-12 by mouse splenic DCs. Mouse splenic DCs were cultured with SAC (20 μg/ml) in the presence of IFN-γ and titrated concentrations of GM-CSF, for 23 h. The supernatants were analyzed for IL-12 p70 and p40 by ELISA (A and B) and Western blotting (C). The result is typical of three experiments with ELISA readouts and Western blotting.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Titration, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: The effects of IL-4 on p70 expression are mediated by the regulation of p35 mRNA. Mouse splenic DCs were cultured with CpG alone or with the addition of IFN-γ, IL-4, or both, for 4 h. RNA was extracted from the cells and subjected to Northern transfer analysis. The relative amounts of radioactive p35-specific or p40-specific probes bound to the mRNA samples as quantitated using a PhosphorImager ® (Molecular Probes) are shown. A repeat experiment confirmed these findings.

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Expressing, Cell Culture, Northern Blot

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: The Influence of IL-4 on the Production of IL-12 p70 and p40 In Vivo

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques:

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: IL-12 p70 and p40 Production by Isolated Human Thymic DCs in the Presence or Absence of IL-4

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques: Isolation

Journal: The Journal of Experimental Medicine

Article Title: Interleukin (Il)-4 Is a Major Regulatory Cytokine Governing Bioactive IL-12 Production by Mouse and Human Dendritic Cells

doi:

Figure Lengend Snippet: The Influence of IL-4, IFN-γ, and GM-CSF on the Production of IL-12 p70 and p40 of Human MoDCs

Article Snippet: In brief, 96-well polyvinyl chloride microtiter plates (Dynatech Laboratories) were coated with the appropriate purified capture mAb, namely R2-9A5 (anti–mouse IL-12 p70; American Type Culture Collection), 20C2 (anti–human IL-12 p70; BD PharMingen), C15.6 (anti–mouse IL-12 p40; BD PharMingen), or C8.3 (anti–human IL-12 p40; BD PharMingen).

Techniques:

Journal: eLife

Article Title: Histone deacetylase 3 represses cholesterol efflux during CD4 + T-cell activation

doi: 10.7554/eLife.70978

Figure Lengend Snippet:

Article Snippet: For T h 2 differentiation, media was supplemented with 1 μg/ml of each anti-IFNγ (XMG1.2, Bio X Cell #BE0055) and anti-IL-12 antibody (R2-9A5, Bio X Cell BE0233) as well 10 ng/ml of IL-4 (PeproTech #214–14).

Techniques: Flow Cytometry, Cell Culture, Blocking Assay, In Vitro, Recombinant, Staining, Isolation, Selection, Sequencing, Software