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. Author manuscript; available in PMC: 2015 Mar 24.
Published in final edited form as: Methods Mol Biol. 2011;699:119–164. doi: 10.1007/978-1-61737-950-5_7

Fig. 7.

Fig. 7

Sorting logic for Teff, Treg, and accessory cells. Monocyte-depleted lymphocytes prepared from TRIMA filters were stained with CD127 PE, CD4 PE-Cy7, and CD25 APC and then sorted into populations of Teff, Treg, and accessory cells to study the effects of Teff and Treg on each other’s proliferative response. (a) Histograms were sequentially gated by applying the lymphocyte scatter region (R1 on FSC vs. SSC plot) and side scatter-based multiplet exclusion criteria (R2 on SSC-H vs. SSC-W plot); R1 and R2 to the SSC-A versus CD4 PECy7-A plot; and R1, R2, and R3 to the CD25 APC-A versus CD127 PE-A plot. Teff cells were defined as CD4+ CD127bright CD25dim singlet lymphocytes. Treg cells were defined as CD4+ CD127dim CD25+ singlet lymphocytes. Accessory cells were defined as CD4− singlet lymphocytes. (b) After sorting, purity was assessed for each population and found to be greater than 97% in all cases.