Extended Data Figure 6. Purification of Human Immune Cell Subsets.

(a) Immune populations subjected to epigenomic profiling in this study (red labels) or prior publications. (b) CD4⁺ cells were enriched based on CD25 expression (MACS) and subsequently sorted based on CD25^highCD127^low/− to isolate T_reg cells; confirmed with FOXP3 intracellular staining. (c) CD4⁺CD25⁻ cells were sorted to isolate T_mem (CD45RO⁺CD45RA⁻) and T_naive (CD45RO⁻CD45RA⁺) cells. (d) CD4+CD25− cells were PMA/ionomycin stimulated and separated based on IL17 surface expression (MACS and FACS) to isolate Th17 cells (IL17⁺) and Th_Stim cells (IL17⁻). (e) Naïve (CD45RA⁺CD45RO⁻) and memory (CD45RA⁻CD45RO⁺) CD8⁺ T cells were isolated using a BD FACSAria 4-way cell sorter. Results are shown from one of two large-scale sorts. (f) Mononuclear cells were isolated from pediatric tonsils. Following CD10 enrichment (MACS), B centroblasts (CD19⁺CD10⁺CXCR4⁺CD44⁻CD3⁻) were purified by FACS.