Extended Data Fig. 8: Ifngr1-deficient T_reg cells from infected mice maintain suppressive function of naïve CD4⁺ T_conv cells in vitro and do not impact T_H polarization of CD4⁺ T_conv cells in vivo.

a,b, In vitro microsuppression assay of splenic uninfected CD4⁺ T_conv (T_responders) by splenic T_reg cells from Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice infected with LCMV Arm. (a) or Cl. 13 (b) (numbers of mice used indicated in parentheses). c, In vitro microsuppression assay of splenic CD44^hiCD62L^lo CD4⁺ T_conv (T_eff) from Arm. infected Foxp3^Cre-YFP mice by splenic T_reg cells from Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice infected with LCMV Arm. (numbers of mice used indicated in parentheses). d-g, Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice infected with Cl. 13 and expression of T-bet, GATA-3 and RORγt (d) (n = 8), T-bet and GATA-3 co-expression (e) (n = 8), CXCR3 (f) (n = 8) and CCR4 (g) (n = 8, control; n = 9, Ifngr1-deficient) by splenic CD4⁺ T_conv cells were measured by flow cytometry. h, Expression of the deleted region of Ifng, normalized to the intact region of Ifng, from Ifng^L/LFoxp3^Cre-YFP mice (n = 4) compared to Foxp3^Cre-YFP mice (n = 3). Thy1.2^– cells, CD8⁺ T cells, CD4⁺ T_conv cells and T_reg cells were sorted, gDNA was isolated and qPCR was performed. a-h Data are presented as mean values and represent biologically independent mice and 1 (h), 2 (b-g) or 3 (a) independent experiments. Statistical significance was determined by two-way ANOVA (a-c), or by multiple unpaired Student t-tests (d, e, h), or by an unpaired two-tailed Student’s t-test (f, g), relative to Foxp3^Cre-YFP mice (P values indicated when significant); NS, not significant.

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