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Coculture of CD25− CD4+ T cells with naive splenic dendritic cells. About 1×106 CD4+ CD25 cells isolated from spleens of naive SJL/J mice were cocultured in 2ml of complete DMEM with CD11c+ dendritic cells also isolated from spleen with or without YFAK, YEAK or LPS in the culture medium. After 7 days, cell surface CD4, CD25 and intracellular Foxp3 were analyzed by FACS.
