T cell activation induces normal proliferation and effector T cell differentiation in the absence of Foxp4.

<p>A) C57BL/6 CD4⁺ T cells were isolated and plated in wells coated with 1 µg/mL anti-CD3 and 5 µg/mL soluble anti-CD28. Wells were harvested on the indicated days. RNA was isolated and cDNA was generated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042273#s2" target="_blank">Materials and Methods</a>. Foxp4 relative to β-actin transcript levels were normalized to day 0. Representative of 2 experiments. B) CD4⁺ T cells were isolated from spleens of cHET and cKO mice, labeled with CFSE, and cultured four days in wells coated with indicated concentrations of anti-CD3 and 5 µg/mL soluble anti-CD28. Dilution of CFSE was assessed on day 4. Representative of 4 cHET and cKO mice. C) CD4⁺ T cells from cHET and cKO mice were stimulated overnight in wells coated with anti-CD3 plus soluble anti-CD28. Harvested cells were stained for CD69 and cytokine receptor IL-7R. Histograms are gated on CD4 or CD8 TCRβ⁺ cells. Representative of 2 experiments. D) cHET and cKO CD4⁺ T cells were cultured in wells coated with anti-CD3 plus soluble anti-CD28. After four days, cells were stained for expression of activation markers CD44 and CD62L. Representative of 4 cHET and 5 cKO mice.