Fig. 1
The expression of FXR in osteogenic differentiation and bone formation. a Schematic graph of the RNA-seq strategy to identify Nuclear Receptor Superfamily genes which may regulated osteogenic differentiation. Total RNA was isolated from BMSCs, which was cultured from 4-week male WT mice, after 4 days of osteoblastic differentiation followed by RNA sequencing analysis. Kolmogorov–Smirnov (K–S) test was used for testing the correlation between the Control and osteoblast differentiation set. b Heatmap of Osteoblast-related genes and Nuclear Receptor Superfamily mRNA expression in BMSCs after 4 days of osteoblastic differentiation. OB Osteoblast differentiation. c–g Quantitative RT-PCR analysis of RUNX2, Sp7, Alp, Col1a, FXR expression in BMSCs after 3 and 7 days of osteoblastic differentiation (n = 3). h Western blot analysis of FXR, RUNX2 and Sp7 during osteoblast differentiation in the mouse BMSCs. Samples were collected at 0, 1, 3, 7, and 10 days after differentiation. i Western blot analysis of FXR in femoral osteoblasts at mice different developmental stages. j, k Representative immunofluorescence images (j) and quantification (k) of FXR (red) positive cells and Osteoblast marker (Sp7, green) on embryonic day 18.5, 1 week, 2 weeks and 4 weeks femur from C57BL/6 J mice. The nuclei were visualized via DAPI (blue) staining. GP means Growth Plate. Scale bar = 100 μm. n = 5. *P < 0.05, **P < 0.01, ***P < 0.001. Data are represented as mean ± SEM
