Fig. 4: T cell intrinsic AR signaling shapes T cell functional populations. | Nature Communications

Fig. 4: T cell intrinsic AR signaling shapes T cell functional populations.

From: Androgens contribute to sex bias of autoimmunity in mice by T cell-intrinsic regulation of Ptpn22 phosphatase expression

Fig. 4

Unbiased clustering of CD4+ (a) and CD8+ T cells (b) by Single Cell RNAseq in the PLNs of 8-week-old NOD.Arf/y or NOD.Arf/yCD4Cre+ male mice. Number of mice: NOD.Arf/y n = 3, NOD.Arf/yCD4Cre+ n = 3. Clustering was performed aggregating CD4+ or CD8+ T cells from all mice using Leiden clustering. Tables show proportions of cells present in each cluster represented as the mean%± SD across the three replicates in each group, weighted by the number of cells the replicate contributes to the aggregated population of CD4+ or CD8+ T cells for that group. P-values were calculated using a χ2 test comparing the distribution of cells within and outside each cluster between Cre− and Cre+ groups. UMAP visualizations depict Ledien clustering as well as clusters enriched in either Cre− (orange) or Cre+ mice (blue). Top differentially expressed genes characterizing significantly different clusters are shown in Supplementary Fig. 2d, e. c–e Flow cytometry analysis of CD4+ (d) and CD8+ (e, f) T cells from PLNs of 10-week-old NOD.Arf/yCD4Cre− (filled bars) and NOD.Arf/yCD4Cre+ (open bars) males. Mean ± sem. Significance was calculated using an unpaired t-test (c–e). Single cell PLN suspensions were stained with mAbs specific for CD4, CD8α, CD44, CD62L, CD69, FoxP3, and Fc block and CD8α, CD44, CD62L, CD25, CD69, IFNγ, TNFα, and Fc block, respectively. Naïve, CD4+ or CD8+, CD62LhiCD44lo; activated, CD4+ or CD8+, CD62LloCD44hi. Number of mice per group, from left to right: (c) n = 7, n = 7, n = 4, n = 4, n = 3, n = 3, n = 7, n = 4, and (d) n = 3, n = 3, n = 4, n = 4, n = 3, n = 3, n = 10, n = 7, n = 3, n = 3, n = 3, n = 3. e Flow cytometric analysis of CD8+ T cells from PLNs of 10-week-old NOD.Arf/yCD4-Cre− (filled bars) and NOD.Arf/yCD4Cre+ (open bars) males. Combined data from two experiments. Number of mice: MPEC, NOD.Arf/yCD4-Cre− n = 3, NOD.Arf/yCD4Cre+ n = 3, SLEC, NOD.Arf/yCD4-Cre− n = 6, NOD.Arf/yCD4Cre+ n = 3. For gating schemes see Supplementary Fig. 5.

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