Fig. 2

HOTAIR overexpression in MSCs delays bone development. a, b Alcian blue and Alizarin red staining of the whole skeletons of WT or Prx1-HOTAIR TG mice at 5 days old. Scale bar, 1 cm. c Representative images showing three-dimensional trabecular architecture by micro-CT reconstruction from WT and Prx1-HOTAIR TG male mice at 1 month old. Scale bar, 0.5 mm. d Representative images showing the three-dimensional trabecular architecture and cortical architecture as shown by micro-CT reconstruction at the distal femurs from WT and Prx1-HOTAIR TG mice at 1 month old. Scale bar, 0.5 mm. e Micro-CT measurements of BV/TV, BMD, Tb.N, Tb.Th, Tb.Sp, and Cort.Th in the distal femurs of mice. n = 10 for each group. BV/TV, ratio of bone volume to tissue volume; BMD, bone mineral density; Tb.N, trabecular number; Tb.Th, trabecular thickness; Tb.Sp, trabecular separation; Cort.Th, cortical bone thickness. f Representative images showing new bone formation assessed by double calcein labeling in WT and Prx1-HOTAIR TG mice. Scale bar, 20 μm. g Quantification of mineral apposition rate (MAR) and osteoblast number to bone surface (N.ob/BS) (n = 6 for each group). h The maximal (max.) load at failure determined by three-point bending of femurs from WT (n = 10) and Prx1-HOTAIR TG (n = 10) mice. i Histological images for Ocn staining of the proximal tibia from WT and Prx1-HOTAIR TG mice. Scale bar, 100 μm. j ELISA analysis of the PINP protein level in the serum from WT (n = 10) and Prx1-HOTAIR TG (n = 10) mice. k qRT-PCR analysis of Alp, Runx2 and Sp7 mRNA levels in bone tissues collected from WT (n = 10) and Prx1-HOTAIR TG (n = 10) mice. Two-tailed unpaired Student’s t test was used for statistical evaluations of two group comparisons. All data are the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001