Fig. 1: Membrane-bound LTα₁β₂ expression increases during thymic T_reg development concomitantly with OX40 and GITR.

a Gating strategy used to analyze conventional CD4⁺ SP thymocytes (CD4⁺ T_conv, CD4⁺CD8⁻Rag2^GFP+CD25⁻Foxp3⁻; black), CD25⁺ T_regP (CD4⁺CD8⁻Rag2^GFP+CD25⁺Foxp3⁻; blue), Foxp3^lo T_regP (CD4⁺CD8⁻Rag2^GFP+CD25⁻Foxp3^lo; green), and mature T_reg (CD4⁺CD8⁻Rag2^GFP+CD25⁺Foxp3⁺; red) in Rag2^GFPxFoxp3^Thy1.1 mice by flow cytometry. b Representative histograms and quantification of membrane-bound LTα₁β₂ expression, detected using the soluble LTβR-Fc receptor, in CD4⁺ T_conv, CD25⁺ T_regP, Foxp3^lo T_regP, and mature T_reg from Rag2^GFPxFoxp3^Thy1.1 mice and Lta^−/− controls. c Geometric MFI (gMFI) of LTα₁β₂ expression in thymic T_reg subsets (n = 10 from three independent experiments. d Correlation between LTα₁β₂ and OX40 or GITR expression in thymic T_reg subsets (n = 9 from three independent experiments). e Gates represent the expression spectrum of CD25 and Foxp3 (left) in mature T_reg that were analyzed for LTα₁β₂ expression (right). The gray histogram corresponds to mature T_reg stained with only the secondary antibody. f Correlation between LTα₁β₂ and CD25 (left) or Foxp3 expression (right) in each gates (n = 10 from three independent experiments). Correlations were calculated using the parametric two-tailed Pearson correlation test for (d) and the non-parametric two-tailed Spearman correlation test for (f). Error bars show mean ± SEM, *p < 0.05, **p < 0.01 and ****p < 0.0001 using one-way ANOVA. Source data are provided as a Source Data File.