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. 2019 Sep 10;9(9):669. doi: 10.3390/ani9090669

Figure 4.

Figure 4

UCP3 is a target of miR-152. (A) Schematic illustration of the predicted binding site for miR-152 in UCP3 3’UTR. The wild-type miRNA-binding sites in UCP3 3’UTR (red) were modified to complementary sequences (blue) to construct the mutated UCP3 3’UTR. (B) The predicted binding site of miR-152 in UCP3 3’UTR is highly conserved among mammals. (C) The sketch map of the dual-luciferase reporter vector psi-CHECK2. The putative miR-152 target site of UCP3 3’UTR and mutation target site were inserted into the psi-CHECK2 vector at the 3’ end of the Renilla luciferase gene (hRluc). The Firefly luciferase activity was treated as an internal normalization control. (D) The luciferase reporter containing the wild type and mutant UCP3 3’UTR was co-transfected into HEK293T cells with miR-152 mimics or mimics control. At 48 h post-transfection, the Renillas luciferase activity was measured and normalized to the firefly luciferase activity. (E) UCP3 protein expression in the LD muscles of LW and RC pigs (n = 5, five pigs), α-Tubulin served as the reference. (F) UCP3 protein expression in different types of skeletal muscle in adult LW pigs (n = 5, five pigs), α-Tubulin served as the reference. Results are presented as mean ± S.E.M. (n = 4, four independent experiments). * p < 0.05, ** p < 0.01 and *** p < 0.001 as contrasted with the control.