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. 2019 Feb 20;10(3):172. doi: 10.1038/s41419-019-1428-1

Fig. 7. BMP/Smad signaling is regulated by microRNA-185 (miR-185).

Fig. 7

a Six weeks after ovariectomized (OVX) operation, the mice femurs were dissected and total RNA was extracted. The expression level of Biglycan (Bgn) and Bmp2 were determined by real-time PCR (n = 3). b The expression of Bgn, Bmp2, t-Smad1, p-Smad1/5/8 in wild-type (WT) or knockout (KO) bones after OVX was detected by western blot. c, d Mesenchymal stem cells (MSCs) were derived from WT or miR-185 KO mice 6 weeks after OVX operation, and transfected with Bgn small interfering (siRNA) or negative control (NC). Cells were cultured with osteoblast induction medium (OIM) for 7 days and alkaline phosphatase (ALP) staining (c) and quantification (d) were carried out. e, f MSCs were derived from OVX mice, and cultured with OIM. Bgn siRNA or NC transfection was carried out at day 0 and day 7. Alizarin Red S staining (e) and mineralization quantification (f) were performed at day 14. g The expressions of Alp, Bgn, Bmp2, t-Smad1, p-Smad1/5/8 in MSCs were detected by western blot after OIM induction for 4 days, and expressed as densitometry normalized to GAPDH. Data were shown as mean ± SD (ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001)