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. 2013 Dec 5;8(12):e82054. doi: 10.1371/journal.pone.0082054

Figure 2. Validation of predicted ssc-miR-24 target genes.

Figure 2

To confirm the legitimacy of selected miR-24 binding sites, a previously developed [REF] in vitro luciferase assay was utilized. Gateway cloning (Life Technologies) was used to create RCAS viruses expressing either the ssc-miR-24 hairpin or a scrambled control (SC) hairpin. DF1 cells, a chicken embryonic fibroblast cell line, were infected with either the RCAS-miR-24 expressing virus or the RCAS-SC expressing virus were transfected with predicted target 3′ UTR luciferase constructs. Target constructs were generated by cloning the miRNA binding site and its flanking regions downstream of the Renilla luciferase gene in the psiCHECK2 vector (Promega). Relative Renilla luciferase activity is shown. *: p<0.05, **: p<0.01. aThis construct contains two miR-24 binding sites. bThis construct contains two miR-24 binding sites. cThis construct contains three miR-24 binding sites.