Figure 1.

Osteogenic differentiation evaluation and MIR99AHG expression quantitation.(a) Relative MIR99AHG expression in senile mice injected with the lentiviruses of shNC or shMIR99AHG and those without injection was quantified via qRT-PCR. β-actin was used as internal reference. (b) Micro-CT analysis on senile mice injected with the lentiviruses of shNC or shMIR99AHG was conducted. (c) Immunohistochemistry on the bone tissue of mice injected with the lentiviruses of shNC or shMIR99AHG was performed. Magnification: × 100. Scale bar = 50 μm. (d) Relative mRNA expression of Osx, Col1A1, OCN, and RUNX2 in BMSCs on day 1, 3, 7 and 14 post osteogenic differentiation was also quantified with qRT-PCR. β-actin was the internal reference. (e-f) Osteogenic differentiation of BMSCs was evaluated by both ALP and Alizarin Red S staining. Magnification: × 200. Scale bar = 100 μm. (g) Relative MIR99AHG expression in BMSCs on day 1, 3, 7 and 14 post osteogenic differentiation induction was further measured with qRT-PCR. β-actin was the internal reference.

All experiments have been performed independently in triplicate and data were expressed as mean ± standard deviation (SD).

^{^^^}p<0.001, vs. shNC; *p<0.05, **p<0.01, ***p<0.001, vs. 0 d.

MIR99AHG: Mir-99a-Let-7c Cluster Host Gene; BMSCs: bone marrow mesenchymal stem cells; OP: osteoporosis; qRT-PCR: quantitative real-time PCR; shRNA: short hairpin RNA; NC: negative control; ALP: Alkaline phosphatase; Osx: Osterix; Col1A1: Collagen, Type I, Alpha 1; OCN: Osteocalcin; RUNX2: RUNX Family Transcription Factor 2.