Figure 3.
γ:δ T cells are not altered in Pten cKO mice. (A) Pten mRNA expression in CD4+ T cells, neutrophils, γ:δ T cells, dendritic cells, and ILC3 cells was measured by qRT-PCR. Each cell type was isolated from WT (Ptenfl/fl) and cKO (Ptenfl/flIl17acre) mice. Data were normalized to Gapdh expression. Data were pooled from three individual experiments. (B) Development of γ:δ T cells in the spleen, pLNs, and thymus was measured by flow cytometry after cells were isolated from WT and cKO mice. (C) Splenocytes from WT and cKO mice were stimulated with PMA/ionomycin for 4 h, and gated on CD3ε+ CD4+ population (top) or CD3ε+ γ:δ TCR+ population (bottom), and IL-17A expression was measured. Data were pooled from three individual experiments (right). (D) Mononuclear cells were isolated from colonic lamina propria. Cells were stimulated with PMA/ionomycin for 4 h, and IL-17A expression by CD4+ T cells was measured by flow cytometry. Data were pooled from three individual experiments (right). Dot plot data in B–D are representative of three individual experiments. Error bars represent the SD. Statistical differences between groups were determined by the Student t test. *, P < 0.05; **, P < 0.01; and ***, P < 0.001.