The small short hair RNA (shRNA) targeting LINC00899 (shLINC00899) with its negative control (shNC), miR-374a mimics with its negative control (NC mimics), and miR-374a inhibitor with its negative control (NC inhibitor) were purchased from GenePharma (Shanghai, China). To overexpress LINC00899, the fulllength LINC00899 sequence was inserted into the pcDNA3.1 vector (GenePharma, Shanghai). Cell transfection was carried out by Lipofectamine 2000 (Invitrogen). The subsequent experiments were performed following 48 h of transfection.
Bioinformatic prediction and dual-luciferase reporter assay Starbase 2.0 (http://starbase.sysu.edu.cn/) was used to predict the binding sites between miR-374a and LINC00899 or RUNX2. The pmirGLO-LINC00899-WT/Mut and pmirGLO-RUNX2-WT/Mut reporters were obtained from GenePharma (Shanghai, China). Then miR-374a mimics or NC mimics was co-transfected with these above reporters into 293T cells. 48 h after transfection, the relative luciferase activity was detected using dual-luciferase reporter assay system (Promega).
Bioinformatic prediction and dual-luciferase reporter assay Starbase 2.0 (http://starbase.sysu.edu.cn/) was used to predict the binding sites between miR-374a and LINC00899 or RUNX2. The pmirGLO-LINC00899-WT/Mut and pmirGLO-RUNX2-WT/Mut reporters were obtained from GenePharma (Shanghai, China). Then miR-374a mimics or NC mimics was co-transfected with these above reporters into 293T cells. 48 h after transfection, the relative luciferase activity was detected using dual-luciferase reporter assay system (Promega).