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. 2019 Jul 17;10:1526. doi: 10.3389/fimmu.2019.01526

Figure 1.

Figure 1

Reciprocal effects of daurinol on Th17 and Treg cell differentiation in murine CD4+ T cells. Splenic CD4+ T cells from DBA/1J mice were cultured under the Th17-inducing condition (cultured with plate-bound anti-CD3 (0.5 μg/ml), soluble anti-CD28 (0.5 μg/ml), anti-interferon-γ anti-IFNγ; [5 μg/ml (A) or 2 μg/ml (D)], anti–IL-4 [5 μg/ml (A) or 2 μg/ml (D)], IL-6 (20 ng/ml), and TGFβ (2 ng/ml) for 72 h in the presence of vehicle or daurinol. Three days later, the cells were stained with antibodies to CD4, IL-17, and FoxP3. (A) A plot from one representative experiment shows the frequencies of IL-17+, CD25+, and FoxP3+ cells among CD+ T cells. (B) Mean ± SD values are presented in the form of a histogram. Data are representative of four independent experiments with similar results. (C) IL-17 levels in culture supernatants shown in (A,B) were measured by ELISA. (D) CD25+, Foxp3+, or IL-17+ cells among CFSE-labeled proliferating CD4+ T cells cultured under the Th17-skewing condition were analyzed by flow cytometry. (E) Expression of Treg-related genes (Foxp3, TGF-β, SOCS3; upper panel) and Th17-related genes (IL-17, RORγt, HIF-1α; lower panel) in the experiments shown in (D) were determined by real-time PCR. Data were obtained from three independent experiments, and values are represented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. –, untreated; Veh, vehicle-treated (DMSO); Dau, daurinol.