Fig. 4

rAAV9-mediated silencing of shn3 prevents bone loss in a mouse model of osteoporosis. a, b Sham or OVX surgery was performed on 3-month-old female Shn3^+/+ and Shn3^−/− mice and 2 months later, femoral trabecular bone mass was assessed by microCT. Representative images of the femur (a) and relative BV/TV (b) are displayed (n = 6 per group). c–j Diagram of the study and treatment methods (c). Sham or OVX surgery was performed on 3-month-old female mice and 6 weeks later, a single dose of 4 × 10¹¹ genome copies of rAAV9 carrying amiR-ctrl or amiR-shn3 was i.v. injected. Seven weeks after injection, femurs were cryo sectioned to identify EGFP-expressing cells (d). shn3 mRNA levels in tibial bone are displayed after normalization to hprt (n = 8–12 per group) (e). Femoral trabecular bone mass was assessed by microCT. Representative 3D reconstruction (f) and relative quantification (g) are displayed (n = 7–8 per group). Representative images of calcein/alizarin red labeling (h) and relative histomorphometric quantification of BFR/BS and MAR (i). Arrows indicate the distance between calcein and alizarin red labeling. Femoral biomechanical properties, including bending rigidity, bending moment, apparent bending modulus, and apparent bending stress were quantified (n = 5–9 per group) (j). Scale bars: a, f 1 mm; d 250 μm; and h 50 μm. Values represent mean ± SD: N.S., not significant; *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001 by an unpaired two-tailed Student’s t-test (b) and one-way ANOVA test (e, g, i, j)