Fig. 5.

Aire deficiency in the thymic stroma impairs the recirculation and the suppressive signature of CCR6⁺ T_reg. A Experimental setup: lethally irradiated CD45.2 Aire^WT or Aire^KO recipients were reconstituted with BM cells from CD45.1 Foxp3^eGFP mice. Six weeks later, thymic T_reg subsets of CD45.1 origin were analyzed by flow cytometry. CCR6⁺ T_reg were cell-sorted to measure the expression levels of genes associated with their suppressive functions. B Flow cytometry profiles, frequencies and numbers of CD4⁺ SP thymocytes of CD45.1 origin in the thymus of BM chimeric mice. C, D Flow cytometry profiles and numbers of CD25⁺ T_regP, Foxp3^lo T_regP and total CD25⁺Foxp3⁺ cells (C) as well as of CCR6⁻ and CCR6⁺ cells in CD25⁺Foxp3⁺ T_reg (D) of CD45.1 origin in the thymus of BM chimeras. Data are derived from 2 independent experiments (n = 4 mice per group and per experiment). E The expression level of Foxp3, Klrg1, Il10, Tgfb1, Gzmb, Fasl, Lag3, Entpd1 and Nt5e was measured by qPCR in purified CCR6⁺CD25⁺Foxp3⁺ T_reg from CD45.1 Foxp3^eGFP → Aire^WT (n = 8) and CD45.1 Foxp3^eGFP → Aire^KO (n = 9) chimeras. Data are derived from 2 independent experiments (n = 4–5 mice per group and per experiment). Bar graphs show mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using unpaired Student’s t test for (B-D) and two-tailed Mann–Whitney test for (E)