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In vitro TH2 cells differentiation
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CD4+CD25−CD62L+ naïve T cells were sorted from C57BL/6 WT mice and differentiated in a TH2-polarizing condition (5 μg ml−1 anti-IFN-γ and 10 ng ml−1 IL-4) using plate-bound α-CD3/α-CD28 and low serum (3–5% FBS)-containing RPMI medium. Afterwards, the resulting cells were re-stimulated in serum free medium (OpTmizerTM CTSTM T-Cell Expansion SFM, Life Technologies) for intracellular cytokine expression, apoptosis and proliferation assay, and siRNA silencing assay in the presence or absence of leptin (200 ng ml−1) as indicated.
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