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. Author manuscript; available in PMC: 2012 Jul 15.
Published in final edited form as: J Immunol. 2011 Jun 17;187(2):692–701. doi: 10.4049/jimmunol.1003658

FIGURE 2.

FIGURE 2

Gamma secretase inhibitors (GSIs) significantly reduce Th17 cytokine levels in human in-vitro Th17 polarization assays. (A) ELISA of IL-17A, IL-17F and IL-22 in supernatants of Th17 polarized naïve human CD4+ T cells treated with GSIs or DMSO as a vehicle control. Purified human CD4+ T cells were pretreated with GSIs (2μM ILCHO and 5μM Compound E) or DMSO as a vehicle control and then cultured in Th0 and Th17 polarizing conditions. Supernatants were collected at 24, 48 and 72 hours and were analyzed for IL-17A, IL-17F and IL-22. (B) Whole cell lysates were prepared from naive CD4+ T cells un-stimulated (US), or differentiated under Th0 and Th17 conditions and immunoblotted for interacellular active Notch1. β-actin was used to confirm equal loading. (C) Naïve CD4+ T cells were activated in-vitro under Th17 polarizing conditions for 4 days, followed by treatment with either DMSO or GSI. Supernatants were collected after 24 h and IL-17A and IL-22 ELISA were performed. Data shown here is representative of three independent experiments done in triplicates. *p≤0.05, ** ≤0.001.