Figure 5.

lncRNA-ORLNC1 Directly Targets miR-296, and miR-296 Possesses the Ability to Regulate Osteogenic Differentiation of BMSCs

(A and B) The effects of miR-106a, miR-106b, miR-20a, miR-20b, miR-296-3p, and miR-346 on the osteogenic differentiation of BMSCs were detected by ARS (A) and ALP (B) stainings. Scale bar, 100 μm. (C) FISH images showing localization of lncRNA-ORLNC1 in BMSCs. 18S, probe for 18S rRNA; U6, probe for U6 snRNA. Scale bar, 200 μm. (D) Luciferase reporter assay was used to assess the relationship between lncRNA-ORLNC1 and miR-296. (E) The expression level of lncRNA-ORLNC1 was upregulated in BMSCs transfected with biotin-labeled miR-296. (F) The expression of miR-296 was significantly decreased in the BMSCs (left) and bone tissue (right) of osteoporotic mice normalized to the control group. (G) The level of miR-296 was markedly decreased in the BMSCs (left) and bone tissue (right) of osteoporotic transgenic mice compared with WT mice. (H) Quantification of miR-296 in the bone tissue of patients with osteoporosis compared with control. (I and J) The expression level of miR-296 was highly increased during osteogenic differentiation (I) and obviously decreased after adipogenic differentiation (J) of BMSCs. **p < 0.01, and ***p < 0.001.