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. 2012 Nov 6;7(11):e48974. doi: 10.1371/journal.pone.0048974

Figure 2. Assessment of knockdown potency using reporter genes.

Figure 2

(A) Schematic drawing of constructed reporter genes. The psiCheck2-backbone reporter plasmid encoding the Renilla luciferase gene carrying the target sequence of miR-29a (psiCheck2-miR29a) or miR-29b (psiCheck2-miR29b) was constructed. The SV40 promoter, Renilla luciferase coding region (CDR) and 3′ untranslated region (3′UTR) of the reporter genes are indicated. The target sequences of miR-29a and miR-29b are indicated in the 3′UTRs. (B, C) Suppression of reporter genes by synthetic miR-29 mimics. The reporter plasmids indicated were transfected together with synthetic miRNA mimics of miR-29a (mimic_29a), miR-29b (mimic_29b) or a negative control miRNA (negative_miR) into N2a cells. 24 hr after transfection, dual-luciferase assay was carried out. The target (Renilla) luciferase activity was normalized to control (Photinus) luciferase activity and further normalized to the data obtained from N2a cells transfected with only the reporter plasmid (non-treatment). Data are averages of four independent experiments. Error bars represent standard deviations. The normalized data were analyzed by one-way analysis of variance (ANOVA), followed by Dunnett's test. Significant differences are indicated by * (P<0.05).