Fig. 4

Hsa_circ_0001485 affected osteogenic differentiation through the TGFβ-BMP pathway in the osteogenic hFOB 1.19 cells. A qPCR determined the knockdown efficiency of three siRNAs targeting BMPR2. B, C Western blot analysis of the protein expression of BMPR2 in osteogenic hFOB 1.19 cells transfected with si-BMPR2 and quantification by densitometric analysis. D AR and ALP staining was performed on the cells transfected with si-BMPR2. For AR, magnification, 100 × , scale bar = 200 μm; for ALP, magnification, 200 × , scale bar = 100 μm. E, F The expression levels of RUNX2, OPN, and OCN in cells transfected with si-BMPR2 were detected using western blot and quantitative analysis. G Osteogenic hFOB 1.19 cells were transfected with OE-circ1485 and then transfected with si-BMPR2 or DMH1 inhibitor, and qPCR measured the expression of hsa_circ_0001485 and BMPR2. H AR and ALP staining was performed on cells transfected with OE-circ1485 and then transfected with si-BMPR2 or DMH1 inhibitor. For AR, magnification, 100 ×, scale bar = 200 μm; for ALP, magnification, 200 ×, scale bar = 100 μm. I, J Osteogenic hFOB 1.19 cells were transfected with OE-circ1485 and then transfected with si-BMPR2 or DMH1 inhibitor. Western blot analysis of the levels of BMPR2, Smad1/5/9, p-Smad1/5/9, Smad4, RUNX2, OPN, and OCN in osteogenic hFOB 1.19 cells and quantification of western blot bands by densitometric analysis. The results are representative data from three replicates, and the data mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001 vs. NC group; ^#P < 0.05; ^##P < 0.01; ^###P < 0.001 vs. OE-circ1485 group)