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. 2014 Apr 15;7(5):2209–2218.

Figure 2.

Figure 2

The expression of Cbl was suppressed by miR-9 and miR-181a. A: Schematic diagram for constructing the predict miR-9 and miR-181a binding sites into pmirGLO vector. B: Cbl is the target gene of miR-9 and miR-181a. RAW264.7 cells were co-transfected with miRNA control, miR-9/181a mimic, anti-miR control or miR-9/181a inhibitor and pmirGLO-Cbl for dual-luciferase assay. When 4 nucleotides of the binding sites of miR-9 or miR-181a in the 3’-UTR of Cbl were mutated (pmirGLO-Cbl-Mu1 and pmirGLO-Cbl-Mu2), the luciferase activities were significantly decreased in RAW264.7 cells co-transfected with miR-9 mimic and wild type Cbl 3’UTR vector and pmirGLO-Cbl-Mu2 compared with pmirGLO-Cbl-Mu1. The luciferase activities were significantly decreased in RAW264.7 cells co-transfected with miR-181a mimic and wild type Cbl 3’UTR vector and pmirGLO-Cbl-Mu1 compared with pmirGLO-Cbl-Mu2. C: Cbl protein level in miRNA mimics or inhibitors-treated RAW264.7 cells was detected by western blot.