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. 2022 Jul 14;13:4075. doi: 10.1038/s41467-022-31722-5

Fig. 2. IL-6 and TGFβ are the primary inducers of Hh signaling in Th17 cells.

Fig. 2

a–c Naïve CD4+ T cells were purified from spleen and peripheral lymph nodes of C57BL/6 mice and polarized with the indicated polarizing cytokines. a, b Cells were harvested at day 3 post stimulation for immunoblot analysis of Smo and qRT-PCR analysis of Gli1 and Gli3. TGFβ blocking antibody (clone: 1D11) was added in the indicated condition for the duration of polarization at 10 μg/ml. Data is normalized to Tbp as a reference gene. Similar results were obtained when CD3ε was used as a reference gene. n = 2–3 independent experiments. c Cells were polarized in vitro under full Th17-polarizing conditions and harvested at day 3 post stimulation for qRT-PCR of Gli3 and Smo. Cells were treated for three days with either 2 μM STATTIC, 1 μM SB 505124, or carrier control; naïve CD4 + T cell RNA levels (“0 h”) are shown as controls. Data is normalized to Tbp as a reference gene. Similar results were obtained when CD3ε was used as a reference gene. n = 4. Data are means +/− SD. p-values were calculated using a two-way ANOVA (a, b) or one-way ANOVA (c) with Tukey's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p< 0.0001 . Source data are provided in the source data file.