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. 2019 May 20;10:1120. doi: 10.3389/fimmu.2019.01120

Figure 2.

Figure 2

ATP is essential for NO-mediated induction of IL-9 in human Th9 cells. (A) Sorted naïve T cells differentiated under Th0 and Th9 polarizing conditions for 6 days in the presence of ATP and Suramin followed by examination of mRNA expression of NOS2 and nitrite production in the culture supernatants. (B) Sorted naïve T cells differentiated under Th0 and Th9 polarizing conditions for 6 days in the presence of NOD (NO donor) and NOI (NO inhibitor) followed by examination of mRNA expression of IL-9, IRF4 and NOS2. (C) Nitrite measurement in the culture supernatants. (D) IL-9 production in the culture supernatants estimated by ELISA (E) Intracellular cytokine staining for IL-9 production. (F) Sorted naïve T cells differentiated under Th0 and Th9 polarizing conditions for 6 days in the presence of NOI (NO inhibitor) and ATP+NOI followed by examination of mRNA expression of IL-9 and NOS2 and ELISA estimation for IL-9 production. (G) Intracellular cytokine staining for IL-9. Data are representative of mean ± SEM from three independent experiments (n = 3). *P < 0.0332, **P < 0.0021, ***P < 0.0002, ****P < 0.0001; one-way ANOVA followed by Tukey's test (A–D, F).