As described previously29 (link), CD4+ T cells were isolated from the spleens of WT and GPR43−/− mice by using anti-mouse CD4-magnetic beads (BD Biosciences). To polarize Th17 cells, CD4+ T cells were cultured with 10 ng/ml TGF-β1, 30 ng/ml IL-6, 10 µg/ml anti-IFN-γ, and 10 µg/ml anti-IL-4 and activated with anti-CD3 and anti-CD28 for 5 d.
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Th17 Cell Polarization from Mouse CD4+ T Cells
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Corresponding Organization : Tongji University
Other organizations : Bristol-Myers Squibb (United States)
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Variable analysis
independent variables
- Mouse genotype (WT vs GPR43-/- mice)
- Th17 cell polarization
- CD4+ T cell isolation using anti-mouse CD4-magnetic beads
- Th17 cell polarization conditions (10 ng/ml TGF-β1, 30 ng/ml IL-6, 10 µg/ml anti-IFN-γ, 10 µg/ml anti-IL-4, anti-CD3 and anti-CD28 activation)
- Positive control: WT mice
- Negative control: GPR43-/- mice
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