Effect of miR-142-3p on expression of endogenous IL-6 protein and mRNA levels in DCs. (A) Knockdown of miR-142-3p de-represses IL-6 expression: IL-6 levels were measured by ELISA in the supernatants from DCs treated with or without LPS and those transfected with either miR-142-3p or miR-223 knockdown or scramble probes for 24 or 48 hours. Data shown are the pooled results from 4 separate experiments (mean ± SEM). (B) IL-6 protein levels are reduced by the overexpression of miR-142-3p: IL-6 was measured by the ELISA in the supernatants of DCs treated with LPS or diluents and ELISA in DCs transfected with either miR-142-3p knockdown or scramble probes or pre-miR-142 duplex or scramble control for 48 hours. Data shown are the pooled results from 3 separate experiments. (C) IL-6 mRNA expression is altered with either knockdown or overexpression of miR-142-3p: IL-6 mRNA levels were analyzed by quantitative RT-PCR in DCs that were transfected with either miR-142-3p knockdown or pre-miR-142 overexpression or scramble controls for 48 hours. Data are combined from 5 experiments with similar results (mean ± SEM). (D) Kinetics of IL-6 mRNA expression following miR-142-3p knockdown and LPS treatment: DCs were transfected with miR-142-3p knockdown or scramble probes for 24 hours, and then treated with LPS for the indicated time. IL-6 mRNA levels were analyzed as in panel C. Data shown are combined results from 3 similar experiments (mean ± SEM).