Fig. 6. Ablation of anti-silencer promotes differentiation of Triplelo Treg subset.
a) FACS analysis of Foxp3 versus CD4 (top row), PD-1 versus GITR (second row), CD4 versus CD25 (third row), or CD25 expression by gated GITRloPD-1lo or GITRhiPD-1hi LN (mesenteric) T cells. Bar graphs at right represent % of Tregs amongst total CD4 lymphocytes (top), or % of Triplelo and Triplehi cells amongst Foxp3+ CD4+ T cells, of ThPOKΔOB11/+ or ThPOKΔOB11/ΔOB11 mice. b) RNA was collected from WT CD4+ Foxp3+T cells subsetted into Triplehi, TripleINT and Triplelo subsets, as indicated, and assayed for ThPOK mRNA expression by qPCR. c) FACS analysis of Foxp3 versus CD25 (top row), PD-1 versus GITR (second row), CD4 versus CD25 (third, fourth rows), and CD73 versus FR4 expression for WT or ThPOKΔOB11/ΔOB11 CD4 LN T cells following in vitro Treg polarization. Plots are for total population or gated subsets, as indicated. d) FACS analysis of Foxp3 versus forward scatter (FSC) (top row), CD4 versus CD8α (second row), and PD-1 versus GITR (bottom row) by total or gated Foxp3+ cLPLs or cIELs T cells, as indicated. Bar graphs at bottom represent % of Tregs amongst total population (left), or % of Triplelo and Triplehi cells amongst gated T cells, of ThPOK +/+ or ThPOKΔOB11/ΔOB11 mice, as indicated. e) FACS analysis of CD4 versus CD8α (top row), or CD8α versus CD8β (second, third rows) expression, for either total TCRβ+ cells or indicated gated subset of freshly isolated iIEL, cIEL, iLPL or cLPL populations, as indicated. i-IEL =intestinal IEL; c-IEL = Colonic IEL; i-LPL = intestinal LPL, c-LPL = colonic -LPL.