Daan Pieren

89 Compromised DNA repair promotes the accumulation of regulatory T cells FSC-A SSC-A 33.3 96.2 96.5 99.1 7.6 5.9 49.9 36.1 CD4 + Mem CD4 + Naive CD4 - Mem CD4 - Naive FSC-A Live/Dead (BV510) FSC-A FSC-H CD3 (FITC) CD44 (BV421) Live/Dead (BV510) CD4 (BUV395) CD44 (BV421) CD4 (BUV395) Lymphocytes Single cells Viable cells Viable CD3 + cells CD4+ and CD4- naive and memory cells Supplementary Figure 2. Gating strategy towards naive and memory CD4 + and CD4 - T cells that were analyzed by viSNE. Representative FACS plots show the gating from spleen lymphocytes to single cells, viable cells, and viable CD3 + cells. These CD3 + T cells were further subdivided into four T-cell subsets: naive CD4 + T cells (CD44 Lo CD4 + CD3 + ), memory CD4 + T cells (CD44 Hi CD4 + CD3 + ), naive CD4 - T cells (Tc cells) (CD44 Lo CD4 - CD3 + ), memory CD4 - T cells (Tc cells) (CD44 Hi CD4 - CD3 + ). Numbers indicate the percentage of events in each gate as part of all events in the FACS plot. The gated naive and memory cell subsets were subsequently analyzed by dimensionality reduction (viSNE). 3