Research Paper Volume 15, Issue 18 pp 9499—9520

Figure 5. SMAD2 knockdown reverses the effect of downregulated miR-486-5p expression on BMMSC differentiation. (A, B) Western blots and qRT-PCR were used to analyse osteogenic factor expression after different treatments. (C) ALP activity assays were performed to analyse ALP activity during osteogenic differentiation on days 0, 3, and 7. (D) ALP staining was performed on day 14, and Alizarin red staining showed increased calcification on day 21 after different transfections. Scale bars, 200 μm. (E, F) Western blotting and qRT-PCR were performed to analyse the protein and mRNA expression levels of adipogenic markers, respectively. (G, H) Oil red O staining and extraction were performed to detect lipid droplet formation on day 10 of adipogenic differentiation. Scale bars, 200 μm. Data are expressed as the mean ± SEM. ^*p < 0.05, ^**p < 0.01, ^***p < 0.005.