Figure 10. HRas activity rescues Tr1 cell development and IRF4 expression in Itk^−/− cells.

WT and Itk^−/− naive IL-10^GFP/Foxp3^RFP reporter CD4⁺ cells were cultured under Tr1 differentiation conditions, retrovirally transduced with control-YFP, WT HRas-YFP or its constitutively active mutant HRas^G12V-YFP. (a) Representative FACS plots of IL-10^GFP+ T cells. (b) Summary of percentage of IL-10^GFP cells over viable CD4⁺Foxp3⁻ T cells and relative IL-10 MFI in IL-10⁺Foxp3⁻ cells (WT control level set as 1). (c) Representative FACS plots of IRF4 expression in RV-transduced cells. Naive CD4⁺ T cells were used as control in grey. n=6. Data represent results of three experiments. *P≤0.05, ***P≤0.001; NS, no significance, by one-way ANOVA with Tukey’s post-hoc test. Data presented as mean±s.e.m.