Figure 2: AU-rich elements within the Ifng 3’UTR determine protein production in mouse and human T cells.
OTI cells were transduced with deletion mutants (a) or with ARE mutants of the full length murine Ifng 3’UTR (c-e). (b) Sequence of the murine Ifng 3’UTR. AREs are underlined. (a,c) Representative GFP levels of resting OTI cells (gray histograms), and after reactivation with OVA257–264 peptide for 6h (black lines). (d) GFP-MFI of resting OTI cells transduced with indicated ARE mutants. (e) Fold increase of GFP-MFI upon activation with OVA257–264 peptide compared to non-activated GFP-expressing T cells. (d-e) Data are presented as mean±SD of at least 3 independently performed experiments [one-way ANOVA with Dunnett’s multiple comparison to the control vector; n=8 mice per group; *p<0.005; **p<0.0005]. (f) Human T cells were transduced with full length human GFP-IFNG 3’UTR (WT), or with ARE mutants. Dot plots depicting GFP-MFI levels from resting T cells (top), or from PMA/ionomycin reactivated T cells (bottom), with CD8+ T cells shown in black and CD4+ T cells in gray. Data shown are representative of 4 individuals, and of 3 independently performed experiments. Compiled data are depicted in Supplementary Fig. 2d.