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. 2013 Oct 1;8(10):e76784. doi: 10.1371/journal.pone.0076784

Figure 9. In vitro target assay for effects of the chicken miR-1689* on expression of the Sp1 transcript.

Figure 9

(A) Diagram of miR-1689* binding site in the 3′-UTR region of the Sp1 transcript. (B) The maps of eGFP-Sp1 3′-UTR expressing and DsRed-miR-1689* expressing vector. The Sp1 3′-UTR sequence was subcloned between the eGFP gene and the polyA tail, generating the fusion construct of the GFP transcript following the Sp1 3′-UTR (left panel). A miRNA expressing vector was designed to generate DsRed and miR-1689* fused transcript or only DsRed transcript for the experimental control (right panel). (C and D) After co-transfection of pcDNA-eGFP-Sp1 3′-UTR and pcDNA-DsRed-miR-1689* into 293FT cells, the fluorescence signals of GFP and DsRed were detected using FACS (C) and fluorescent microscopy (D). In the panel C, a gated event value from histogram statistics was used to find statistical percentages of the positive events (GFP-expressing cells) and the numbers indicate mean±SEM of three independent experiments.