Daan Pieren

54 Chapter 2 SUPPLEMENTARY FIGURES Young (2 months) Aged (28 months) CD4 CD8 4.9 64.7 29.3 6.8 49.5 42.6 CD3 + cells SSC-A Live/Dead FSC-A FSC-H CD3 Live/Dead CD4 FoxP3 Lymphocytes Single cells Viable cells Viable CD3 + T cells T cell subsets Young (2 months) Aged (28 months) 94.0 84.0 92.6 88.0 94.4 76.5 32.5 28.0 0.1 7.3 16.9 0.9 46.1 36.1 41.4 51.2 Treg Th Tc Treg Th Tc A B 0 2 4 6 8 10 % CD4 - CD8 - of CD3 + cells Young (2 months) Aged (28 months) ns Supplementary Figure 1. Gating strategy towards Th, Tc, and Treg cell subsets and the pro- portion of CD4 - CD8 - T cells in young and aged mice. ( A ) Plots show the gating strategy from splenocyte lymphocytes, single cells, viable cells, viable CD3 + cells towards the three T cell subsets; Th cells (FoxP3 - CD4 + of CD3 + ), Tc cells (FoxP3 - CD4 - of CD3 + ), and Treg cells (Foxp3 + CD4 + of CD3 + ) in a young (2 months old) and an aged (28 months old) mouse representing their age group. ( B ) Plots and graphs show the proportion of CD3 + T cells that do not express CD4 and CD8 in young and aged mice. Mean ± SD; ns = not statistically significant for difference between young and aged mice using Mann-Whitney test.