Fig. 4.

Regulatory and memory type T cells in Bcl10^–/– in Pkcθ^–/– mice. (A and B) FACS analysis of CD4⁺CD25⁺ T_R cells in the thymus (A) and of T_R and memory type T cells in the spleen (B) of Bcl10^–/–, Pkcθ^–/–, and appropriate control mice. Numbers indicate mean and SD of the percentages of CD4⁺CD25⁺ T cells of CD4-SP thymocytes (A) and of CD25⁺CD45Rb^int T and CD25^–CD45Rb^low T cells of splenic CD4 T cells (B), determined from five mice, respectively. Pkcθ^–/– mice were on a pure C57BL/6 genetic background, whereas the Bcl10^–/– mice were on a C57BL/6–129 mixed genetic background; therefore, the data obtained from the respective age- and sex-matched control mice are shown separately. (C) RT-PCR showing levels of Foxp3- and Hprt-specific transcripts amplified from total mRNA purified from CD4 and CD8 T cells from Bcl10^–/–, Pkcθ^–/–, and control mice. One of at least three independent experiments is shown for each genotype. (D) Quantification of Foxp3 mRNA levels in indicated T cell subsets purified from WT (n = 6), Pkcθ^–/– (n = 3), and Bcl10^–/– (n = 3) mice by real-time PCR. Foxp3 mRNA expression is normalized to expression of Hprt mRNA: Foxp3_Hprt-message. The mean Foxp3_Hprt-message in CD4 T cells purified from WT mice was set to 1. Foxp3_Hprt-message in WT CD8 T cells and CD4 and CD8 T cells purified from Pkcθ^–/– and Bcl10^–/– mice was calculated relative to Foxp3_Hprt-message (WT CD4 T cells) = 1. Results are shown as the mean, and error bars indicate the SEM.