Figure 4.

miR-672-5p Treatment Represses Smurf1 In Vivo to Promote Osteoblastogenesis

(A) ALP activity (OD, n = 6), (B) mineralization (OD, n = 6), and (C) representative wells of Cfu-ob formation. Data are mean ± SE. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the PBS-treated group, or as shown ˆˆp < 0.01. mRNA expression (qPCR, in triplicate) of Smurf1 (D), Runx2 (E), and BMP2 (F) and protein expression (G) (western blotting) of Smurf1 (ab38866-Abcam; 1:1,000), Runx2 (ab76956-Abcam; 1:1,000), and BMP2 (ab14933-Abcam; 1:1,000) in osteoblasts from sham-operated (Sham) or ovariectomized (OVx) mice that were given PBS (0.2 mL), scrambled miR (miR-C, 7.0 mg kg⁻¹), or miR-672-5p (7.0 mg kg⁻¹). β-actin (sc-47778 Santa Cruz Biotechnology; 1:500) was taken as loading control. Secondary antibodies (either anti-rabbit or anti-mouse; 1:10,000) were HRP conjugated (Sigma-Aldrich). Data are mean ± SE. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the PBS-treated group. See also Figures S2D–S2F for in vitro actions of microRNA-672-5p on osteoclastogenesis from bone marrow cell cultures.