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186710.fig.002b
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Figure 2: Plasmin induces time- and concentration-dependent expression of CCL20 mRNA in dendritic cells. (a) Plasmin does not induce release of the proinflammatory cytokines TNF-α, IL-1α, and IL-1β. At day 6, dendritic cells were left untreated or stimulated with plasmin (0.143 CTA U/mL) for 24 h. Release of cytokines in the culture media was analyzed with the proteome profiler array. (b) Plasmin induces time-dependent expression of CCL20 mRNA in dendritic cells. Dendritic cells were treated with plasmin (0.143 CTA U/mL) or the positive control LPS (0.5 μg/mL) for the indicated time. mRNA was isolated and subjected to RT-PCR (b) and real time qPCR (c) analysis using CCL20-specific primers; GAPDH served as control. Results are mean ± SEM of 3 experiments. (d) Dendritic cells were treated for 6 h with the indicated concentrations of plasmin, and CCL20 mRNA expression was analyzed by RT-PCR. (e) Proteolytic activity of plasmin is required for the induction of CCL20 mRNA expression. Dendritic cells were stimulated either with plasmin (0.143 CTA U/mL) or the equivalent amount of catalytically inactivated plasmin (VPLCK plasmin) for 6 h. CCL20 mRNA expression was analyzed by RT-PCR. All data are representative of at least 3 independent experiments.