Fig. 5
M2D-Exos induce osteoblastic differentiation of BMSCs in vitro. a miRNA-5106 was upregulated in agomiR-5106 treated BMSCs following transfection using lipofecamine control, 200 µM agomiR-5106, agomiR-NC, antagomiR-5106 or antagomiR-NC for 48 h; b Western blotting analysis of Collagen I, ALP, OCN and Runx2 protein levels in BMSCs treated using lipofecamine control, agomiR-NC, agomiR-5106, antagomiR-NC or antagomiR-5106 for 48 h; c The relative intensity of the western blotting analysis; d qRT-PCR analysis was used to assess expression of osteoblast differentiation genes including Col1a1, ALP, OCN, and Runx2 in transfected BMSCs; e M2D-Exos were treated with miR-5106 inhibitor (antagomiR-5106) to investigate the effect of M2D-Exos with miR-5106-silence on the osteoblastic differentiation of BMSCs. The results showed that antagomiR-5106 can significantly decrease the miR-5106 expression in M2D-Exos; f Decreased osteogenic genes expression can be detected in M2D-ExosantagomiR−5106 group compared with PBS and M2D-Exos groups; g Alizarin red-mediated calcium staining in BMSCs following differently transfeced for 21 days, Scale bar = 10 mm; h ALP staining in BMSCs following different treatments for 14 days. Scale bar = 10 mm. Data are mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001