miR-134 downregulates ITGB1 by directly targeting its 3′UTR.

<p><b>A.</b> Schema of the candidate genes identified by four independent approaches. <b>B.</b> Putative binding sequences of ITGB1 for miR-134 in the human (Hsa), chimpanzee (Ptr), rhesus (Mml) and mouse (Mmu). The ITGB1 seed sequences are highlighted by ruby italic letters. <b>C.</b> Putative miR-134-binding sites in the wild-type ITGB1 3′-UTR (highlighted by ruby letters) or 3′-UTR segments containing mutant (highlighted by blue letters) binding sites are shown. <b>D.</b> Relative luciferase activity assays for constructs containing luciferase and wild-type or mutant ITGB1 3′-UTRs were performed after cotransfection with a miR-134 mimic or negative control. The <i>Renilla</i> luciferase vector was cotransfected as an internal control. Data were analyzed by one-way ANOVA (p<0.01), and differences between two groups were assessed by the Bonferroni post-test. <b>E.</b> The mRNA expression levels of ITGB1 in SK-HEP-1 and Huh-7 cells overexpressing miR-134 or negative control. Differences between two groups were analyzed by the Mann-Whitney test. <b>F.</b> The protein levels of ITGB1 in SK-HEP-1 and Huh-7 cells infected with miR-134 or control lentivirus. GAPDH served as an internal control. <b>G.</b> Protein levels of ITGB1 in Huh-7 cells after transfection with the miR-134 inhibitor or inhibitor control. GAPDH served as an internal control.