Figure 1.

The G-protein-coupled estrogen receptor (GPER) -directed agonist G-1 induces interleukin-10 (IL-10) production from CD4⁺ T cells. CD4⁺ CD44^lo CD62L^hi naive CD4⁺ T cells were collected by FACS and cultured for 4 days ex vivo with various combinations of transforming growth factor-β (TGF-β), IL-6 and IL-23, and supplemented with 100 nm G-1 or vehicle (DMSO, control). Cells were subsequently stained for intracellular IFN-γ, IL-17A and IL-10, then analysed by flow cytometry. (a) Representative plots from the various conditions showing intracellular IL-17A and IL-10. (b) Quantification of data from five to seven independent experiments showing relative number of total IL-10⁺ cells and total IL-17A⁺ cells in cultures treated with TGF-β + IL-6. (c) Quantification of data from four to seven independent experiments showing relative prevalence of total IL-10⁺ cells and total IL-17A⁺ cells in cultures treated with TGF-β + IL-6 + IL-23. (d) Quantification of IFN-γ⁺ cells in cultures stimulated with anti-CD3/28 in non-polarizing conditions (i.e. without the addition of any cytokines). P-values determined by Student's t-test; *P < 0·05; ***P < 0·0005. Error bars = SEM; NS, not significant.