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. 2021 Jan 11;11:609186. doi: 10.3389/fendo.2020.609186

Figure 6.

Figure 6

miR-145a inhibited osteogenic differentiation in C3H10T1/2 cells by targeting Cbfb. The potential miR-145a target genes predicted by TargetScan and miRDB (A). Putative miR-145a binding site in Cbfb 3’-UTR (B). The mRNA level (C) and protein level (D) of Cbfb measured respectively by qRT-PCR and Western blot in C3H10T1/2 overexpressed miR-145a. Luciferase reporter assay of Cbfb 3’-UTR (E) (Mean±SD, n=3 per group, the luciferase activity was normalized by firefly luciferase. * p < 0.05). Western blot analysis of the expression of Runx2 and Cbfb in C3H10T1/2 cells transfected with miR-145a mimic and pcDNA3.1-Cbfb and then cultured in osteogenic differentiation medium (ODM) for 3 days (F) and Alkaline phosphatase (ALP) staining (G) and Alizarin Red staining (H) was carried out for these cells on the 7th, 21th day, respectively. Results of mRNA qRT-PCR were normalized to GAPDH expression. Results are presented as the mean±SD (*p < 0.05; **p < 0.01; ***p < 0.001). Bar: 200μm.