Figure 3. Foxp4 is not required for the generation of peripheral CD4 and CD8 T cells.

A) Absolute numbers from spleens from four-week-old cHET and cKO mice, including total spleen cellularity (top panel), numbers of CD4⁺ T cells (middle panel) and CD8+ T cells (bottom panel). B–C) Splenocytes cells from 6–8 week old cHET and cKO mice were stained for expression of CD4, CD8 and CD44 and CD62L or CD25, CD69, TCRβ, and IL-7 receptor (CD127) for immunophenotyping by polychromatic flow cytometry on LSRII. Histograms are gated on live, singlet CD4⁺ T cells. Representative of 4 independent experiments. D) Splenic Foxp3⁺ nTreg cells were identified by flow cytometry based on expression of CD4, CD25 and intracellular staining for Foxp3 protein. Gated frequencies are indicated. Representative of 7 cHET and 7 cKO mice. E) Frequencies of Treg were calculated using the splenic cellularity and relative frequency. Each point represents an individual mouse. Mean and standard deviation are indicated. F) Naïve CD4 T cells from cHET or cKO mice were polarized for four days in vitro in the presence of IL-2 with (left) or without (right) TGFβ. Wells were harvested and cells were assessed for expression of CD25 and Foxp3. Plots are gated on live, CD4⁺ cells. Representative of 4 experiments.