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. 2016 Feb 26;11(2):e0149600. doi: 10.1371/journal.pone.0149600

Fig 1. CDDO-Me redirects LPS-induced cytokine production of human M2 macrophages.

Fig 1

Human peripheral blood-derived monoctyes were differentiated with M-CSF to generate alternatively activated (M2-polarized) macrophages. (A) CDDO-Me dose-dependently induced TNF-α secretion in LPS-stimulated macrophages. Culture supernatants from macrophages treated with CDDO-Me or DMSO control for 16 hrs prior to stimulation with LPS (10 ng/ml) for 24 hrs were analyzed by ELISA for TNF-α. Data are representative of results obtained with 3 individual donors, and depict average values obtained from 3 independent assays. (B) ELISA analysis of IL-6, IL-10, TNF-α and VEGF secretion from human macrophages pretreated with 300 nM CDDO-Me for 16 hrs followed by activation with LPS as in (A). CDDO-Me effects on IL-6 secretion were assayed in 5 donors; 3 donors were assayed for effects on TNF, IL-10, and VEGF. (C) Total RNA was extracted from macrophages pretreated with or without CDDO-Me (300 nM) for 16 hrs followed by LPS for 24 hrs. mRNA transcript levels were measured by Taqman real time PCR *p<0.05, **p<0.01, ***p<0.005, ****p<0.001 vs. untreated control. Data shown are representative of results obtained from analysis of 3 individual donors. CDDO-Me effects on mRNA levels were analyzed in 3 separate experiments for each donor. As indicated in Methods, 3 technical replicates were analyzed in each separate, individual experiment.