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. 2018 Dec 7;27(2):394–410. doi: 10.1016/j.ymthe.2018.11.019

Figure 2.

Figure 2

lncRNA-ORLNC1 Inhibits Osteogenesis and Stimulates Adipogenesis of BMSCs

(A) ARS staining and quantification deriving from osteogenic BMSCs with lentivirus overexpressing lncRNA-ORLNC1 or lncRNA-NC were performed to assess the osteogenic capacity after 14 days. Cells grown in the normal growth medium were shown as a reference. NM, normal growth medium; OM, osteogenic induced medium. Scale bar, 100 μm. (B) BMSCs stained with ALP staining showed calcium deposits in the presence of lncRNA-ORLNC1 overexpression compared with NC. Scale bar, 100 μm. (C) The expression levels of ALP, Bmp4, Bglap, Spp1, and Col1a1 significantly decreased after treatment with overexpression of lncRNA-ORLNC1. (D) Immunostaining of BMSCs transfected with lncRNA-ORLNC1 overexpression exhibited decreased Runx2-positive cells. Runx2, red; DAPI, blue. Scale bar, 100 μm. (E) Relative protein levels of Runx2 and Osterix after treatment with lncRNA-ORLNC1 overexpression or lncRNA-NC at day 7 of the osteogenic differentiation. (F) Increase of adipogenic differentiation by lncRNA-ORLNC1 overexpression treatment. Scale bar, 200 μm. (G) The mRNA levels of adipogenic genes. (H) Images showed Pparg expression in BMSCs treated with lncRNA-ORLNC1 overexpression compared to NC. Pparg, green; DAPI, blue. Scale bar, 100 μm. (I) Western blot was applied to ensure the protein levels of Pparg and Fabp4 in BMSCs. (J) H&E staining and quantification of bone regeneration. Relative bone growth surface area compared to total surface. The black arrows indicate the bone formation, and the red arrows indicate hydroxyapatite. Scale bar, 50 μm. **p < 0.01, and ***p < 0.001.