Figure 8.

lncRNA-ORLNC1 Regulates Osteogenesis and Adipogenesis of BMSCs through Targeting miR-296, and Pten Is a Candidate Target Gene for miR-296

(A) ARS and ALP stainings indicated that lncRNA-ORLNC1 counteracted the increased effect of miR-296 on osteogenic differentiation of BMSCs. Scale bar, 100 μm. (B and C) Immunofluorescence staining (B) and western blot analysis (C) of relative protein expression showed increased levels of osteogenic genes in miR-296-treated BMSCs were inhibited by lncRNA-ORLNC1. Runx2, red; DAPI, blue. Scale bar, 100 μm. (D) Quantitative analysis of the number of adipocytes revealed that lncRNA-ORLNC1 increased the adipogenic differentiation, which was inhibited by miR-296. Scale bar, 200 μm. (E and F) Immunofluorescence staining (E) and western blot (F) indicated the protein levels of adipocyte-related genes in BMSCs after treatment with lncRNA-ORLNC1 and miR-296. Pparg, green; DAPI, blue. Scale bar, 100 μm. (G) The binding sites between miR-296-3p and Pten. (H) Luciferase reporter assay revealed the relationship between miR-296 and Pten. (I) The relative mRNA level of Pten was not changed after treatment with miR-296 mimic and NC. (J) The protein level of Pten was largely decreased by miR-296 mimic, while it was increased by miR-296 AMO; and, lncRNA-ORLNC1 and miR-296 mimic regulated the protein expression of Pten. *p < 0.05, **p < 0.01, and ***p < 0.001.