Fig. 2.

Developing CCR6⁻ and recirculating CCR6⁺ T_reg are reduced in Aire^KO mice throughout life. A Flow cytometry profiles, frequencies and numbers of CD25⁺ T_regP, Foxp3^lo T_regP and CD25⁺Foxp3⁺ T_reg analyzed in CD4⁺ SP thymocytes from the thymus of 5-day-old Aire^WT and Aire^KO mice. The data are derived from 3 independent experiments (n = 2–4 mice per group and per experiment). B, C Frequencies and numbers of CD25⁺ T_regP, Foxp3^lo T_regP and CD25⁺Foxp3⁺ T_reg in the thymus of 6-week- (B) and 1-year- (C) old Aire^WT and Aire^KO mice. D, E Flow cytometry profiles (D), frequencies and numbers (E) of CCR6⁻ and CCR6⁺ cells in CD25⁺Foxp3⁺ T_reg from 6-week- and 1-year-old Aire^WT and Aire^KO mice. The data are derived from at least 3 independent experiments (n = 3–4 mice per group and per experiment). F The expression of Ccl20 and Cxcl12 was measured by qPCR in purified mTEC^hi (EpCAM⁺UEA-1⁺Ly51^−/loCD80^hi) from 6-week-old Aire^WT and Aire^KO mice. Bar graphs show ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using unpaired Student’s t test (A, B, C, E) and two-tailed Mann–Whitney test for (F)