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. 2020 Aug 26;12(9):2422. doi: 10.3390/cancers12092422

Figure 2.

Figure 2

Multi-parametric Tregs and Tregs subtyping gate-strategy. (a) (i–vi) Representative fluorescence-activated cell sorting (FACS) plots: (i) dead cells identified as positive Live/Dead Horizon V450 and (ii) doublets cells were excluded; (iii) T lymphocytes were gated on the basis of their scatter parameters and (iv) CD4+ T cells were identified and analysed for CD25 and CD127 surface expression: (v) CD4+ T cells expressing high levels of CD25, negative or low expression of CD127, (vi) and FoxP3 positivity was considered as Treg cells (g–h) Treg subpopulation were identified according to Foxp3 level and CD45RA in eTregs (FoxP3hiCD45RA) (b), nTregs(FoxP3intCD45RA+) cells (c) and non-Treg intCD45RA- cells (d).