Fig. 4: Hematopoietic origin of Lta expression regulates thymic T_reg development.

a Experimental set up: lethally irradiated CD45.1/2 WT recipients were reconstituted with CD45.2 WT + CD45.1 Foxp3^eGFP or Foxp3^eGFPxLta^−/− (ratio 1:1) mixed BM cells. Six weeks later, CD45.1 CCR6⁻ T_reg were analyzed in the thymus of recipient mice. b Flow cytometry profiles and frequencies of CD25⁺ T_regP, Foxp3^lo T_regP and mature T_reg from CD45.1 Foxp3^eGFP (n = 12) or Foxp3^eGFPxLta^−/− (n = 12) BM cells. Data are pooled from four independent experiments. c Representative flow cytometry profiles and frequencies of CD25⁺ T_regP, Foxp3^lo T_regP, and mature T_reg in the T_reg cell lineage gate derived from CD45.1 Foxp3^eGFP (n = 6) or Foxp3^eGFPxLta^−/− (n = 7) BM cells. Data are pooled from two independent experiments. d Frequencies of Ki67⁺ cells in CD25⁺ T_regP (n = 9 for Foxp3^eGFP and n = 10 for Foxp3^eGFPxLta^−/−), Foxp3^lo T_regP (n = 10) and mature T_reg (n = 10) derived from CD45.1 Foxp3^eGFP or Foxp3^eGFPxLta^−/− BM cells. Data are pooled from three independent experiments. e-h t-SNE dimensional reduction of flow cytometry data using maturation markers (e) allowing the identification of T_reg clusters (f), fluorescence intensity heatmap of the markers used for the t-SNE construction of each T_reg cluster (g), and quantification of these clusters (h) in cells derived from CD45.1 Foxp3^eGFP (n = 4) or Foxp3^eGFPxLta^−/− (n = 5) BM cells. Data are pooled from two independent experiments. Error bars show mean ± SEM, *p < 0.05, **p < 0.01, and ****p < 0.0001 using unpaired two-tailed Student’s t test for (b) and unpaired two-tailed Mann–Whitney U test for (c), (d) and (h). Source data are provided as a Source Data File.