Fig. 3

miR-381-3p inhibited osteogenic differentiation and bone formation. A,B Expression levels of miR-381-3p in 55-69 and 70-95 years old male (left) and female (right) patients with osteoporosis, as detected by RT-PCR (mean ± S.D., n > 13). C Correlation analysis between miR-381-3p level and CTD-2555A7.2 level in bone tissues from osteoporosis patients, as detected by RT-PCR.D ALP and Alizarin Red staining of hMSCs treated with mimic-381-3p, as detected by ALP staining and Alizarin Red staining (left), and Alp/Runx2 expression levels of hMSCs treated with mimic-381-3p, as detected by RT-PCR (right, mean ± S.D., n = 3). NC: mimic-NC. mimic-381: mimic-381-3p. E ALP and Alizarin Red staining (left), and Alp/Runx2 expression levels (right) of hMSCs treated with inhibitor-381-3p (mean ± S.D., n = 3). iNC: inhibitor-NC. inhibit-381: inhibitor-381-3p. F ALP and RUNX2 protein levels of hMSCs treated with mimic-381-3p, as detected by western blot. Representative western blots visualized by enhanced chemiluminescence method (left) and the quantification results with GAPDH as internal control (right, mean ± S.D., n = 4).G ALP and RUNX2 protein levels of hMSCs treated with inhibitor-381-3p (mean ± S.D., n = 4).H,I Calvarial bone mineral apposition rate of C57BL/6 mice treated with mimic-381-3p (left) or inhibitor-381-3p (right, mean ± S.D., n > 5). Scale bar: 10μm.J Expression and quantification of OCN in calvarial tissues of C57BL/6 mice treated with mimic-381-3p, as detected by immunohistochemical staining (left). Quantification of relative integrated optical density (IOD) values of OCN immunostaining using Image-Pro Plus 6.0 software (right, mean ± SD, n = 3). Scale bar: 25μm.K Expression and quantification of OCN in calvarial tissues of C57BL/6 mice treated with inhibitor-381-3p (mean ± SD, n = 3).*P < 0.05, **P < 0.01, ***P < 0.001