miR-142-3p does not affect the endogenous expression of other cytokine responses by the DCs. (A) Knockdown of either miR-142-3p and miR-223 does not affect other proinflammatory cytokines: levels of TNFα and IL-12 were analyzed by ELISA in the supernatants of DCs treated with LPS or diluents and transfected with either miR-142-3p knockdown or scramble probes or miR-223 knockdown probes for 48 hours. Data shown are combined results from 3 similar experiments (mean ± SEM). (B) The expression of cytokines TNFα, IL-12, and IL-10 are not affected by either the overexpression or knockdown of miR-142-3p: TNFα, IL-12, and IL-10 analyzed by ELISA in the supernatants of DCs treated with LPS or diluent and transfected with either miR-142-3p knockdown or scramble probes or pre-miR-142 duplex for 48 hours. Data shown are combined results from 3 similar experiments (mean ± SEM). (C) miR-142-3p has no direct impact on the expression of other immune molecules: the mRNA levels of TGFBR1, IRAK1, PCAF, and TIRAP were analyzed by quantitative RT-PCR in the DCs that were transfected with either miR-142-3p knockdown or pre-miR-142 duplex or scramble control for 48 hours. Data are combined from 3 experiments with similar results (mean ± SEM). (D) Knockdown of miR-142-3p in DCs does not affect their ability to stimulate allogeneic T cells: allogeneic (BALB/c) T cells were cultured with B6 BM DCs that were transfected with either miR-142-3p knockdown or scramble probes. T-cell proliferation was evaluated at 96 hours following pulsing with ³H for the last 18 hours. Data shown are results from 1 of 2 similar experiments (mean ± SEM).