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. 2012 Nov 7;7(11):e48958. doi: 10.1371/journal.pone.0048958

Figure 2. miR-550a downregulates CPEB4 expression by directly targeting its 3′UTR.

Figure 2

(A) Scheme for the identification of candidate genes combining microarray assays and a target prediction algorithm. (B) The putative binding sequences for miR-550a within the human (Has), chimpanzee (Ptr), rhesus (MMl) and mouse (Mmu) CPEB4 3′UTR. Seed sequences are indicated as emphasized with shadow. (C) The sequences of the putative miR-550a-binding site in the wild-type (emphasized with shadow) and mutant (underlined) CPEB4 3′UTR. (D) The relative luciferase activity of luciferase reporters with wild-type or mutant CPEB4 3′UTRs was determined in HEK 293T cells, which were co-transfected with the miR-550a mimic or negative control (nc). Renilla luciferase activity was analyzed as an internal control. There is statistical significance between the wild-type group and the mutant group with miR-550a transfection. Representative experiments are shown with the mean ± SD. (E) The protein levels of CPEB4 were determined by western blot assays in Huh-7 cells infected with miR-550a or mock vector and Huh-7 cells transfected with the miR-550a inhibitor or negative control (nc).