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. Author manuscript; available in PMC: 2010 May 1.

Published in final edited form as: Nat Immunol. 2009 Sep 20;10(11):1170–1177. doi: 10.1038/ni.1795

Characterization of T_reg cells from Cbfb^fl/flFoxp3^YFP-Cre mice. (a) Flow cytometry of Foxp3 expression in CD4⁺CD8⁻ T cells derived from thymus, LN and spleen of Cbfb^fl/flFoxp3^YFP-Cre (red lines) and littermate Cbfb^fl/+Foxp3^YFP-Cre (blue lines) mice. Numbers over bracketed lines indicate percentages of Foxp3⁺ cells, with the mean fluorescence intensity (MFI) indicated in parentheses. Data represent analyses of at least six different mice in each group. (b) Real-time PCR analyses of Foxp3 mRNA normalized against HPRT1 mRNA in sorted CD4⁺YFP⁻CD62L^hi (T_eff) or CD4⁺YFP⁺ (T_reg) cells from the indicated mice. Data are shown as averages of three independent experiments. (c) Expression of activation markers on peripheral CD4⁺Foxp3⁺ T_reg cells from Cbfb^fl/flFoxp3^YFP-Cre (red lines) and littermate Cbfb^fl/+Foxp3^YFP-Cre (blue lines) mice. The numbers indicate mean and standard deviations and represent at least three independent experiments. Representative CD4⁺Foxp3⁺ gates are shown in the extreme left panel. (d) CD4⁺YFP⁺ T cells sorted from a pool of LNs and spleens of the Cbfb^fl/flFoxp3^YFP⁻ and Cbfb^fl/+Foxp3^YFP-Cre mice were co-cultured with CD4⁺YFP^- responder T cells from wild-type mice at the ratios indicated for 72 hours in the presence of anti-CD3 and irradiated T cell-depleted splenocytes. The data are shown as mean [³H]-thymidine incorporation in triplicate cultures and represent one of two independent experiments. (e) Random X chromosome inactivation in Cbfb^fl/flFoxp3^YFP-Cre/+ mice results in YFP+ and YFP⁻ populations of T_reg cells. Plots show gated CD4⁺ T cells in Cbfb^fl/flFoxp3^YFP-Cre/+ and Cbfb^fl/+Foxp3^YFP-Cre/+ littermates. (f) Bar graphs representing mean frequencies of YFP⁺Foxp3⁺ cells in indicated groups of mice. Data represent three mice per group.