Endogenous miR-142-3p directly targets IL-6 3′ UTRs in DCs. (A) Schematic of the psiCHECK-2 constructs inserted with WT or truncated fragments of IL-6 3′UTR with the distinct miR target sites. (B) Differential regulatory repressions of psiCHECK-2 reporter carrying WT IL-6 3′UTR by endogenous miRNAs in DCs treated with or without LPS. Data shown are representative of results from 3 independent experiments (mean ± SEM). (C) Significantly de-repressed Luc expressions following LPS treatment. Fold changes of expression levels of WT reporters were calculated after first normalizing to empty vector control and were then compared between DCs treated with or without LPS. Data shown are representative of results from 3 separate experiments (mean ± SEM). (D) Differentially suppressed expression ratios of Luc reporters carrying WT or the various truncated IL-6 3′UTR that are mutated at various target sites for the endogenous miRs. Data are representative of 5 separate experiments (mean ± SEM). (E) Differentially regulated de-repression of Luc reporters by LPS treatment. Fold changes were calculated after normalized to empty vector first then compared between DCs treated with or without LPS. Data were obtained over 4 independent experiments (mean ± SEM). (F) Endogenous miR-142-3p predominantly targets IL-6 3′ UTR. Expression levels of Luc reporters carrying WT IL-6 3′UTR were measured after knockdown of either miR-142-3p or miR-223 and after overexpression of miR-142 in DCs treated with or without LPS. The results are from 4 separate experiments (mean ± SEM).