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. 2014 Jul 16;9(7):e102259. doi: 10.1371/journal.pone.0102259

Figure 5. Exploiting endogenous miRNAs to modify exogenous gene expression.

Figure 5

(A) Illustration that miR-125a-5p was selectively reduced in the lymphocytic cell lines, Jurkat and Raji. (B) Schematic of reporter constructs used to assess effect of endogenous levels of miR-125a-5p. Constructs were engineered to contain 1, 2 or 4 miR-125a-5p binding sites or scrambled sequence controls. (C) Meg-01 cells were transfected with the indicated constructs. Luciferase repression was enhanced with more miR-125a-5p binding sites. (D) Meg-01, Raji, Jurkat and K562 cells were transfected with the 4xSC and 4x125 constructs. Luciferase was quantified and normalized to GFP for transfection efficiency. Data plotted as fold-expression compared to constructs with scrambled sequence. (E) Meg-01 cells were co-transfected with Luc-4x125 construct and control locked nucleic acid (LNA) or LNA that specifically inhibits miR-125a-5p. Jurkat cells were co-transfected with Luc-4x125 construct and control pre-miRNA or pre-miR-125a-5p. Data in panels C-E are mean ± SD of at least three independent experiments with two replicates each.