Research Paper Volume 15, Issue 18 pp 9499—9520

Figure 3. miR-486-5p directly enhances osteogenesis but suppresses adipogenesis of BMMSCs. (A) qRT-PCR analysis of miR-486-5p levels in BMMSCs after treatment with mimic-NC, miR-486-5p mimic, inhibitor-NC, or miR-486-5p inhibitor for 48 h. (B) Western blot analysis of COL1, RUNX2, ALP, and BMP in BMMSCs after treatment with mimic-NC, miR-486-5p mimic, inhibitor-NC, or miR-486-5p inhibitor for 48 h. (C) qRT-PCR analysis of the changes in the mRNA levels of the osteogenic differentiation marker genes ALP, OCN, OPN, RUNX2, and COL1 in BMMSCs after treatment with mimic-NC, miR-486-5p mimic, inhibitor-NC, or miR-486-5p inhibitor for 48 h. (D) Relative ALP activity was analysed during osteogenesis in treated BMMSCs on days 0, 3, and 7. (E) ALP staining in BMMSCs following different treatments for 14 days. Scale bars, 200 μm. (F) Alizarin red staining in BMMSCs after different transfections for 21 days. Scale bars, 200 μm. (G, H) The expression of adipogenic-specific markers was analysed by qRT-PCR and Western blots. (I, J) Oil red O staining and extraction were performed to detect lipid droplet formation on day 10. Scale bars, 200 μm. Data are expressed as the mean ± SEM, ^*p < 0.05, ^**p < 0.01, ^***p < 0.005.