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. 2021 Jan 13;20(2):e13298. doi: 10.1111/acel.13298

FIGURE 5.

FIGURE 5

miR‐103‐3p inhibits bone formation in vivo. (a) Live animal fluorescence imaging analysis of GFP expression at 1–3 weeks postinjection (n = 3). (b) Real‐time PCR analysis of miR‐103‐3p levels in bone specimens from mice at 1–3 weeks after treatment with agomir‐103‐3p (n = 3). (c) Real‐time PCR analysis of miR‐103‐3p levels in bone specimens from mice after treatment with agomir‐103‐3p or its negative control. (d) Real‐time PCR analysis of miR‐103‐3p levels in different tissues after a single injection of agomir‐103‐3p, antagomir‐103‐3p, or the corresponding negative controls (n = 3). The relative miRNA levels in the liver tissue were normalized to the mean value of the control group. (e) Schematic diagram illustrating the experimental design. (f) Real‐time PCR analysis of Alp (left), Bglap (middle), and Col1α1 (right) mRNA levels in bone specimens from mice after treatment with agomir‐103‐3p or its negative control. (g) Representative images showing the three‐dimensional trabecular architecture in distal femurs determined by microCT reconstruction. Scale bars, 1 mm. (h) MicroCT measurements of BMD, BV/TV, Tb.Th, Tb.N, and Conn.D in the distal femurs of mice after treatment with agomir‐103‐3p or its negative control. (i) Representative images of new bone formation assessed by double calcein labeling. Scale bars, 10 μm. (j) Double calcein labeling analysis of dynamic bone histomorphometric parameters (MAR, BFR/BS, Ob.s/BS, and Ob.N/B.Pm) in the distal femur of mice after treatment with agomir‐103‐3p or its negative control. (k) Masson's trichrome staining analysis of osteoid formation in the distal femur of mice after treatment with agomir‐103‐3p or its negative control (n = 8). Scale bars, 10 μm. Arrows indicate osteoids. (l) Three‐point bending test analysis of mouse femur biomechanical properties after treatment with agomir‐103‐3p or its negative control. The n value for each group is indicated at the bottom of each bar in the graphs. All data are presented as the mean ± SD. *p < 0.05, **p < 0.01. One‐way ANOVA with a post hoc test was performed, and the significance of differences between two groups was determined with Student's t test