Daan Pieren

55 Novel features of ageing in murine T cells CD4 + T cells CD8 + T cells IFN- γ TNF- α IL-4 + IL-5 Young Aged Medium Medium PMA/ionomycin PMA/ionomycin TNF- α IFN- γ CD4 + T cells CD8 + T cells A B Medium PMA/iono Medium PMA/iono 0 20 40 60 80 IFN- γ + CD4 + T cells (%) *** *** Medium PMA/iono Medium PMA/iono 0 20 40 60 80 TNF- α + CD4 + T cells (%) *** *** *** Medium PMA/iono Medium PMA/iono 0 2 4 6 8 10 IL-4 + IL-5 + CD4 + T cells (%) ** ** Medium PMA/iono Medium PMA/iono 0 20 40 60 80 100 IFN- γ + CD8 + T cells (%) *** *** *** Medium PMA/iono Medium PMA/iono 0 20 40 60 80 TNF- α + CD8 + T cells (%) *** *** *** Medium PMA/iono Medium PMA/iono 0 1 2 3 4 IL-4 + IL-5 + CD8 + T cells (%) * Young (2 months) Aged (28 months) Supplementary Figure 2. Flow cytometry of intracellular cytokines in CD4 + and CD8 + T cells of young and aged mice. ( A ) Plots show the intracellular expression of IFN- γ and TNF- α in CD4 + and CD8 + T cells of a young (2 months old) and an aged (28 months old) mouse representative of their age group after stimulation with PMA/ionomycin or medium as unstimulated control for four hours. ( B ) Graphs show the proportions of IFN- γ + , TNF- α + , or IL-4 + + IL-5 + in CD4 + and CD8 + T cells of young (n=6, 2 months old) and aged (n=4, 28 months old) mice. Mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 for difference between young and aged mice using Two-way ANOVA. 2