(a) CD4+ TN cells were cultured for 3 days with varying concentrations of CD3 (0.1-10.0 μg/ml) and CD28 antibodies (0.5-15.0 μg/ml). Cells were rested for 2 days before activation for 30 min with 20ng/ml IL-6. Intracellular flow cytometry for pY-STAT1 and pY-STAT3 is presented as a fold change in MFI (n=4). (b) Real-time PCR for Ahr, Bcl3, Bcl6, Il10, Il21, Kat2b, Pim1 and Stat3 in CD4+ TN and TEM cells after 30 min stimulation with 20 ng/ml IL-6 (n=3). (c) Hierarchical clustering, using the complete linkage method (row 1-spearman rank correlation, of all significant transcriptomic data (P<0.05, relative signal intensity of >150, and a >1.5 fold) in each CD4+ T cells population with and without treatment with 20ng/ml IL-6 for 6h. (d) Relative number of genes (up-regulated in red and down-regulated in blue), individual fold change and gene intensity expression for genes differentially regulated by IL-6 in CD4+ TN, TEM and TEXP cells (see Supplementary Fig. 2a) at 6h after stimulation in the presence or absence of CD3/CD28 antibodies. Analysis was confined to genes displaying both a relative signal intensity of >150 and >1.5-fold alteration in expression following IL-6 treatment (P<0.05). n.s – not significant. (One-way ANOVA (a). Data are shown as mean ± s.d).