Fig. 5
AAV9-mediated gene silencing of Pum2 alleviates the osteoporotic phenotypes in OVX-induced mice. A Schematic of the time course used for the in vivo experiments in mice with OVX-induced osteoporotic phenotypes. Intravenous injection of 200 μL each of AAV9-NC (approximately 2.5 × 1010 GC/mL) or AAV9-siPum2 (approximately 2.6 × 1010 GC/mlL was administered to commercially available female OVX mice or sham-operated control mice of the same age. Twelve weeks after surgery, the mice were sacrificed for further experiments. B Femoral samples from each group were collected and subjected to μCT analysis to examine osteoporotic phenotypes. Representative μCT images of fracture calluses at twelve weeks post AAV9 injection. μCT showed that systemic injection of AAV9-siPum2 could alleviate bone loss. C The bar graphs shows quantitative analysis of bone-related parameters, including BV/TV, Tb.Th (mm2), Tb.N (1/mm2), and Tb. Sp (mm2) (n = 6). The data are expressed as the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001. BV/TV, bone volume per tissue volume; Tb.Th, trabecular thickness; Tb.N, trabecular number; Tb. Sp, trabecular spacing. D Histological analysis of bone loss and lipid accumulation in the femur was performed using hematoxylin and eosin staining. Scale bar = 1 mm (upper) or 50 μm (lower). Accumulation of lipid droplets in the bone marrow was quantified by counting droplets per mm2 (experimental triplicate, n = 3). The data are expressed as the mean ± SD; ***p < 0.001