Fig. 3

Neat1 deficiency resulted in reduced bone formation and impaired osteoblast function. a, b Representative trabecular and cortical bone 3D μ-CT images from distal femurs of 2-month-old WT and Neat1-KO male mice. Scale bars, 0.5 mm. Cortical bone thickness of distal femurs from the WT (n = 7) and Neat1-KO (n = 7) mice was analyzed. c Quantitative µ-CT analysis of BMD, BV/TV, Tb.N, Tb.Sp and Tb.Th from the WT (n = 7) and Neat1-KO mice (n = 7). d Femur maximal load determined by three-point bending in the WT (n = 6) and Neat1-KO (n = 6) mice. e Double calcein labeling images showing new bone formation from the WT (n = 3) and Neat1-KO (n = 3) mice. BFR/BS, bone formation rate/bone surface. MAR, mineral apposition rate. Scale bars, 50 μm. f Histomorphometric analysis of osteoblast surface per bone surface in the WT and Neat1-KO mice. n = 3 in each group. g Immunohistochemical staining of OCN and Col1α1 in tibial sections from 6-week-old Neat1-KO (n = 3) mice and WT (n = 3) mice. Scale bars, 100 μm. h Analysis of osteogenic marker gene expression in bone tissue from the WT (n = 6) and Neat1-KO (n = 6) mice. i ELISA analysis of serum PINP (ng·mL⁻¹) in 2-month-old WT (n = 6) and Neat1-KO (n = 6) mice. j Representative FISH images of paraspeckles in the WT and Neat1-KO osteoblasts. Scale bars, 10 μm. k Analysis of osteogenic marker genes in primary osteoblasts isolated from the WT and Neat1-KO mice. n = 3 in each group. l ALP staining images of the WT and Neat1-KO osteoblasts induced with osteogenic medium for 3 days. m. Representative Alizarin red staining images of the WT and Neat1-KO osteoblasts. *P < 0.05, **P < 0.01, ***P < 0.001