WTI triggers a local and systemic accumulation of FoxP3+ T regulatory cells. C57BL/6 mice received 0 Gy or 15 Gy whole thorax irradiation. At indicated time points, immune cells were isolated from lung tissue, spleen and cervical lymph nodes (cLN) and stained for flow cytometric analysis. (A) Gating strategy for the detection of FoxP3 on CD4+ T cells. (B) Treg (CD4+ FoxP3+) cells change in the lung during pneumopathy. Mean values ± SEM of the expression of FoxP3 are shown as percentages from CD4+ lung cells. (C) RT-PCR mRNA analysis of FoxP3 expression levels of total lung RNA isolates derived from control as well as whole thorax irradiated animals 21 days after irradiation. βActin was included as control. Two images per condition are shown. (D/E) Timelines of FoxP3 on gated CD4+ T cells in the cLN (D) and spleen (E) during pneumopathy. Shown are mean values ± SEM of percentages calculated on CD4+ LNC and TSC. Cells of 6-9 mice/group were analyzed; ** p ≤ 0.01, *** p ≤ 0.001, two-way ANOVA followed by post-hoc Bonferroni test.