Fig 2. Effect of 25D on differentiation and/or stimulation of monocyte-derived DCs.

Monocytes were differentiated for six days using rGM-CSF and rIL-4 in media with 10% FCS in the presence (25D^diff-DCs or 25D^diff/stim-DCs) or absence (serum-DCs or 25D^stim-DCs) of additional 25D (10⁻⁷ M). Subsequently, 25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (25D^diff/stim-DCs or 25D^stim-DCs) or absence (serum-DCs or 25D^diff-DCs) of additional 25D (10⁻⁷ M), and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig 1 and Fig. 2 were conducted using cells from the same cell preparations of identical donors. *p<0.05, **p<0.01, ^#p≤0.05