Fig. 3: Foxp3 degradation in mature T_reg cells induces expression changes in a small set of genes.

a, T_reg cells from the scRNA-seq dataset were classified as resting or activated based on exceeding the threshold for resting or activated gene signature scores and were subsequently analyzed. b, Scatter-plot showing the correlation of gene expression changes induced by Foxp3 degradation at day 3 and day 7 in resting and activated T_reg cells. FC, fold change. c, UMAP visualization of resting and activated T_reg cells colored by gene signature scores for the ‘TIR1-up’ and ‘TIR1-down’ gene sets, up- and downregulated upon Foxp3 degradation, respectively. d, Dot plot summarizing statistically significant DEGs in resting or activated T_reg cells following 3 or 7 days of 5-ph-IAA-induced Foxp3 degradation. The color represents the log₂ fold change of R26^TIR1(F74G) versus R26^WT and the size represents the Benjamini–Hochberg adjusted P value of the differential expression test. e, Flow cytometry analysis of Foxp3 protein and mRNA levels (reported by ZsGreen) in Foxp3^AID T_reg cells from heterozygous Foxp3^AID/WTR26^WT and Foxp3^AID/WTR26^TIR1(F74G) females after 7 days of Foxp3 degradation. Scatter-plots represent mean ± s.e.m. Data are pooled from two independent experiments and analyzed using a two-way ANOVA. pLN, peripheral lymph nodes. f, Flow cytometry analysis of CD25, CD122, OX40, GITR and FR4 protein levels in Foxp3^AID T_reg cells from heterozygous Foxp3^AID/WTR26^WT and Foxp3^AID/WTR26^TIR1(F74G) females after 7 days of Foxp3 degradation. g, Flow cytometry analysis of CD127 and TCF1 protein levels in Foxp3^AID T_reg cells from heterozygous Foxp3^AID/WTR26^WT and Foxp3^AID/WTR26^TIR1(F74G) females after 7 days of Foxp3 degradation. Scatter-plots represent mean ± s.e.m. (f,g). Each point represents a unique mouse. Data are pooled from two independent experiments and analyzed with two-sided multiple t-tests.

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