Research Paper Volume 12, Issue 12 pp 11754—11767

miR-107 inhibition upregulates CAB39 and activates AMPK-Nrf2 signaling to protect osteoblasts from dexamethasone-induced oxidative injury and cytotoxicity

Figure 1. miR-107 targets and silences CAB39 in osteoblasts. The bioinformatics analyses show that miR-107 putatively targets 3’-UTR of human CAB39 (at position of 1322-1329) (A). The RNA-Pull down assay confirmed the binding between the biotinylated-miR-107 and CAB39 mRNA (normalized to the input control) (B). Stable OB-6 cells with pre-miRNA-107 lentivirus (LV-pre-miR-107-sL1/sL2, two stable cell lines) or non-sense microRNA control lentivirus (“miR-C”, same for all Figures), as well as the parental control OB-6 cells (“Pare”, same for all Figures), were cultured, expression of miRNA-107 and CAB39 was tested by qPCR (C and E) and Western blotting (F) assays, with relative CAB39 3’-UTR luciferase activity (D) examined as well. OB-6 cells were transfected with 500 nM of the applied miR-107 mimics (sequences listed in G) for 48h, CAB39 3’-UTR luciferase activity (H) and its expression (I and J) were tested. The primary human osteoblasts were infected with pre-miRNA-107 lentivirus (LV-pre-miR-107) or miR-C, after 48h expression of listed genes was shown (KM). Data were mean ± standard deviation (SD, n=5). “Trans” stands for the transfection reagent control (HJ). * p<0.05 vs. “miR-C”/“Trans” cells. Each experiment was repeated three times and similar results were obtained.