Figure 1. Rorγ, encoded by Rorc, is preferentially expressed in colonic Tregs.

Gene expression profiles from purified Treg cells of various origins. A: Transcripts that are enriched in tissue and colonic Tregs. Top: transcripts differentially represented in tissue vs. splenic Tregs (at a FoldChange>2). Bottom: transcripts that are most biased in colonic Tregs (FoldChange>1.5 vs any other tissue Treg). Means of at least 2 duplicates. B: Transcription factors overrepresented in colonic Tregs vs other tissue and lymphoid organ Tregs. C: Representative flow cytometry plots of CD4+ T cells, and compilation of frequencies (right) of Rorγ⁺Helios⁻ Tregs within the FoxP3⁺CD4⁺TCRβ⁺ population. Each point is an individual mouse. Data representative of > 3 independent experiments. D: Representative Rorγ vs Helios, Nrp1, Il33R or Gata3 plots for colon or spleen Foxp3⁺CD4⁺TCRβ⁺ Tregs (see Fig. S2 for quantification). Each point is an individual mouse. Data representative of ≥ 3 independent experiments. E: Frequencies of Rorγ⁺Helios⁻ Tregs among FoxP3⁺CD4⁺TCRβ⁺ cells of different tissues (SI: small intestinal lamina propria; PP: Peyer's patches; MLN: mesenteric lymph nodes; scLN: subcutaneous lymph nodes). Each point is an individual mouse. Data pooled from at least 2 independent experiments. F: Flow cytometry analysis of human colon biopsies and frequencies of human RORγ⁺ Tregs within the FOXP3⁺CD4⁺CD8⁻CD3⁺CD45⁺ population. Healthy tissue were endoscopically-determined normal areas from chronic constipation or irritable bowel syndrome patients; inflamed tissue from Crohn's lesions. Each point is an individual patient. Data pooled from 5 independent experiments). G: Il17a (PMA+ionomycin activation, intracellular staining) or Il17f (reporter in Il17f^rfp mice) expression among Foxp3⁺ Treg or FoxP3⁻ Tconv Each point is an individual mouse. Data representative of independent 3 experiments.