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. 2015 Oct 5;10(10):e0139456. doi: 10.1371/journal.pone.0139456

Fig 3. MicroRNA–25 downregulates SOCS4 expression by directly targeting its 3′UTR.

Fig 3

(A) The sequences of miR–25 binding sites within the human SOCS4 3′UTRs and the schematic reporter constructs. In this panel, WT represents the reporter constructs containing the entire 3′UTR sequences of SOCS4. SOCS4-MUT represents the reporter constructs containing mutated nucleotides. (B, C) K1 cells were co-transfected with either WT or MUT SOCS4 3′UTR reporter plasmids and either miR–25 mimic (B) or miR–25 inhibitor (C). Luciferase activities were measured after 48 h using a dual-luciferase assay kit and normalized to Renilla luciferase. (D) K1 cells were transfected with miR–25 mimic or controls for 48 h. SOCS4 mRNA (left panel) and protein (right panel) were detected by qRT-PCR and western blot, respectively. (E) Cells were transfected with miR–25 inhibitor or control and the experiments were performed as in D. (F, G) WRO cells were utilized and experiments were performed as in D and E. All the experiments were repeated at least 3 times with similar results. Bar graphs represent mean ± SD, n = 3 (**P < 0.01; *P < 0.05).