Fig. 1: Key Hedgehog signaling components are induced in Th17 cells.

a Overview of canonical Hh signaling. b–e Naïve CD4⁺ T cells were purified from spleen and peripheral lymph nodes of C57BL/6 mice and stimulated with plate-bound anti-CD3ε/CD28 antibodies in the presence of polarizing cytokines to generate Th0, Th1, Th2, Th17, and iTreg subsets. b Expression of Ihh was assessed by qRT-PCR in naïve CD4⁺ T cells and Th17 cells at the indicated timepoints after TCR stimulation (left) and in mouse embryonic fibroblasts (right). Data is normalized to Tbp as a reference gene. n = 3 independent experiments. c Immunoblot analysis of Ptch and Smo in naïve CD4⁺ T cells and T helper (Th) subsets at indicated timepoints post stimulation. n = 2 (naïve) or 3 (Th subsets) independent experiments. d Immuno-fluorescence imaging (single x-y confocal section) of Th17 cells at day 3 labeled with antibodies against Smo (green). Nuclei were stained with Hoechst (blue). Scale bars: 10 µm. e Expression of Gli1, Gli2, and Gli3 were assessed by qRT-PCR in Th subsets at indicated timepoints post stimulation in the presence of polarizing cytokines. Data is normalized to Tbp as a reference gene with similar results obtained when using CD3ε as a reference gene. n = 3 independent experiments. Gli2 mRNA was undetectable in all conditions tested (see also Supplementary Fig. 2a, b). Source data are provided in the source data file.