Figure 1. ITK is required for Tr1 cell development in vivo.

WT and Itk^−/− IL-10^GFP/Foxp3^RFP dual reporter mice were treated with αCD3ε antibody or control, and live cells from the indicated organs were analysed. (a) Representative FACS plots showing IL-10^GFP expression by Foxp3^RFP− CD4⁺ T cells and summary of IL-10⁺ cell percentage over Foxp3⁻CD4⁺ T cells and number in blood (number per ml), spleen, lung, small intestine and fat (number per gram). (b) Representative FACS plots showing the LAG3/CD49b expression by IL-10^GFP+ Foxp3^RFP− CD4⁺ T cells, summary of LAG3⁺CD49b⁺ cell percentage over IL-10⁺Foxp3⁻CD4⁺ T cells and number of LAG3⁺CD49b⁺ IL-10⁺Foxp3⁻ Tr1 cells from samples shown in (a). ^§P≤0.05, ^§§P≤0.01, ^§§§P≤0.001, compared with the levels in PBS-treated group; *P≤0.05, **P≤0.01, ***P≤0.001, comparing groups connected, by non-parametric Mann–Whitney test. Data were pooled from three experiments, ‘n’ indicates number of replicates in each group/point. Data presented as mean±s.e.m.