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. 2017 Nov 25;9(12):360. doi: 10.3390/v9120360

Figure 2.

Figure 2

cfa-miR-143 targets the Igfbp5 gene. (A) A mutation in the 3′-UTR of the Igfbp5 gene was designed in a seed sequence site for binding with cfa-miR-143. The inserted mutation is marked (red); (B) The WT (wildtype) and mut (mutation) genes were inserted into a dual fluorescence reporter retroviral vector (psicheck-2). The insertion was ligated between the human Renilla luciferase (hRlu) gene and the promoter of the human firefly luciferase (hFlu) gene. The recombined plasmids, psicheck2-Igfbp5 3′UTR WT and psicheck2-Igfbp5 3′UTR mut, were transfected into 293T and MDCK cells along with cfa-miR-143 mimic or mimic NC (Negative Control); (C,D) Detection of relative luciferase activity showed that the hRlu/hFlu ratio was significantly lower (p < 0.05) in cells co-transfected with mimic and WT than in those with mimic and mut in both 293T and MDCK cells. Differences were considered statistically significant at p < 0.05. HSV-TK: herpesvirus thymidine kinase. Differences in expression levels were considered significant if p < 0.05 (* p < 0.05).