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. Author manuscript; available in PMC: 2021 May 3.
Published in final edited form as: J Allergy Clin Immunol. 2018 Aug 7;143(3):1119–1130.e3. doi: 10.1016/j.jaci.2018.06.046

Fig. 3. Blimp-1 expression is not induced during Th9-differentiation.

Fig. 3.

(A-B) Expression of IL-9 and Blimp-1 mRNA in naïve CD4+T cells from WT mice and stimulated with anti-CD3 and anti-CD28 under neutral (medium alone), Th1 (IL-12 and IL-2 plus anti-IL-4), Th2 (IL-4 and IL-2 plus anti-IFN-γ), Th9 (TGF-β and IL-4 plus anti-IFN-γ) and Th9 plus OX86 (TGF-β, IL-4, OX86 and anti-IFN-γ). The expression levels of the indicated genes were analyzed by qPCR. (C-E) Naïve CD4+ T cells from WT mice were cultured in different conditions: stimulation of TCR alone or in the presence of IL-4, TGF-β and OX86 or in combination as shown in Fig. 3E. On day 4, the RNA and culture supernatant were collected, the expression levels of (C) Blimp-1 and (D) IL-9 mRNA were analyzed by qPCR, and (E) the IL-9 levels were measured by ELISA. (F-G) CD4+ T cells were cultured with IL-12+IL-2 (Th1 conditions) for 3 days. In some wells, IL-4+TGF-β+OX86 (Th9 condition) was added along with IL-2+IL-12 (Th1+Th9) on the same day or after 3 days (Th1-3d/Th9) under Th1 conditions. Th1 or Th9 differentiated cells were used as positive or negative controls. On day 6, the RNA and culture supernatants were collected, the expression of (F) Blimp-1 mRNA was analyzed by qPCR, and (G) the IL-9 levels were measured by ELISA. Data (mean ± SD) are representative of three experiments. * P < 0.05; & P < 0.05 and # P < 0.05.