Overexpression of S6K1 rescued the inhibitory effect of ghrelin on Th17 cells.
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Abstract
<p>Total T cells were isolated from mouse spleens and induced differentiation to Th17 cells. Adenovirus was used to infect activated T cells to overexpress S6K1, the critical downstream molecule in mTOR signaling pathway. (A) The phosphorylation of S6 was analyzed with Western Blot. Relative protein signal intensity was quantified. (B&C) Differentiated Th17 cells were treated with adenovirus expressing S6K1 (Ad-S6K1) or GFP (Ad-GFP) with final titer indicated. The mRNA level of RORγt (B) and IL-17A (C) were analyzed with RT-PCR. (D) The concentration of IL-17A in the supernatant was examined with ELISA. (E&F) The percentage of IL-17A<sup>+</sup> cells in splenic total T cells (E) and CD4<sup>+</sup> T cells (F) was analyzed with flow cytometry. Shown is the representative of three independent experiments. *<i>P</i><0.05 versus control; <sup>#</sup><i>P</i><0.05 versus ghrelin-treated alone.</p