Daan Pieren

59 Novel features of ageing in murine T cells A B C 0 3 6 Marker Expression CD3 CD4 FoxP3 CD69 CD25 CD122 PD-1 CTLA-4 GARP CellTrace CD3 CD4 FoxP3 CD69 CD25 CD122 PD-1 CTLA-4 GARP CellTrace CD3 CD4 FoxP3 CD69 CD25 CD122 PD-1 CTLA-4 GARP CellTrace 1 2 3 4 5 6 1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 1 2 3 4 5 6 7 8 9 7 10 11 12 13 1 2 3 4 5 6 8 9 10 1 2 3 4 5 6 7 7 8 9 Day 0 Day 2 Day 4 Day 0 Day 2 Day 4 Day 2 Day 4 Day 0 Day 0 Day 2 Day 4 Day 2 Day 4 Day 0 Day 2 Day 4 Day 0 Young Aged Helper T cells Young Aged Young Aged Cytotoxic T cells Regulatory T cells Supplementary Figure 7. Heatmaps of indicated markers included in viSNE analyses per- formed on Th, Tc, and Treg cells of young and aged mice before and after stimulation. Expression of T-cell markers before and after exposure of splenocytes of young (n=6, 2 months old) and aged (n=4, 28 months old) mice to an intermediate stimulatory strength for two and four days. Heatmaps depict the ArcSinh5-transformed median expression of the designated markers in Th ( A ), Tc ( B ), and Treg ( C ) cell clusters at the indicated days of young and aged mice. Numbers above each heatmap correspond to the cluster numbers identified by viSNE as shown in Figure 6. Red arrows indicate the non-activated (CD25 - CD69 - ) PD-1 + Th and Tc cell clusters in aged mice ( A,B ). Blue arrows indicate the activated (CD25 + CD69 + ) PD-1 + Th and Tc cell clusters in aged mice ( A,B ). 2