miR-27a modulates the expression of <i>RARA</i>, <i>RXRA</i> and <i>PHB</i> genes.

<p><b>A)</b> 293T cells transfected with miR-27a precursors or negative controls (pre-miR-27a or pre-control) analyzed 48h post-transfection were used to perform Luciferase assays in cells transfected with reporter constructs containing the wild type for the indicated genes or a synthetic sequence including a perfect miR-27a binding site (sensor). Results from three independent experiments are shown as mean ± IC of Firefly luciferase activity relative to controls, normalized on Renilla luciferase activity. *P<0.05; **P<0.01; ***P<0.001. <b>B</b>) <i>RARA</i> mRNA levels measured by qRT–PCR in miR-27a over-expressing RH36 cells (pre-miR-27a, light gray box) and negative controls (pre-control, dark gray box) analyzed 24h post-transfection. <b>C</b>) Reporter constructs containing miR-27a wild type or mutant binding sites RARA 3’-UTRs (RARA mut1 and RARA mut2). <b>D</b>) 293T cells transfected with miR-27a precursors (pre-miR-27a or pre-control) analyzed 48h post-transfection were used to perform Luciferase assays in cells transfected with reporter constructs containing miR-27a wild type (RARA) or mutant binding sites in RARA 3’-UTRs (RARA mut1, RARA mut2 and RARA mut1/2). Results from three independent experiments are shown as mean ± IC of Firefly luciferase activity relative to controls, normalized on Renilla luciferase activity. NS = not significant; *P<0.05; **P<0.01; ***P<0.001 (IC = confidence interval). <b>E</b>) <i>RXRA</i> mRNA levels measured by qRT–PCR in miR-27a over-expressing RH36 cells (pre-miR-27a, light gray box) and negative controls (pre-control, dark gray box) analyzed 24h post-transfection. <b>F</b>) Reporter constructs containing miR-27a wild type (RXRA) or mutant binding sites in RXRA 3’-UTRs (RXRA mut). <b>G</b>) 293T cells transfected with miR-27a precursors (pre-miR-27a or pre-control) analyzed 48h post-transfection were used to perform Luciferase assays in cells transfected with reporter constructs containing miR-27a wild type (RXRA) or mutant binding sites in RXRA 3’-UTRs (RXRA mut). Results from three independent experiments are shown as mean ± IC of Firefly luciferase activity relative to controls, normalized on Renilla luciferase activity. NS, not significant; *P<0.05; **P<0.01; ***P<0.001