Introduction
Materials and methods
Experimental animals and analysis of skeletal phenotype
Cell culture
Micro-computed tomography (µCT) analysis
Hematoxylin and eosin (HE) and saffron solid green staining and immunohistochemistry
Immunofluorescence staining
High-throughput quantitative molecular measurement
CCK-8 assay
Western blotting
Statistical analysis
Results
Enpp1 deletion inhibited body weight gain and decreased trabecular bone mass
WT | KO | p value | |
---|---|---|---|
Parameters of µCT | |||
BV/TV (%) | 6.967 ± 0.256 | 1.287 ± 0.220 | < 0.0001 |
Tb. Th(mm) | 0.043 ± 0.003 | 0.037 ± 0.001 | 0.0441 |
Tb. N(mm−1) | 1.651 ± 0.078 | 0.372 ± 0.020 | < 0.0001 |
Tb. Sp(mm) | 0.255 ± 0.009 | 0.449 ± 0.042 | 0.0014 |
Connectivity | 231.333 ± 11.060 | 104.000 ± 14.422 | 0.0003 |
Tb. Pf(mm−1) | 33.477 ± 0.950 | 60.404 ± 2.033 | < 0.0001 |
SMI | 2.259 ± 0.111 | 2.703 ± 0.123 | 0.0096 |
Enpp1 deletion causes reduced proliferation and osteogenesis and increased apoptosis of cells in bone
High-throughput protein sequencing shows downregulated MAPK signaling in the Enpp1 deleted bone
Description | Count | % | Log10(P) | Log10(q) |
---|---|---|---|---|
MAPK signaling pathway | 20 | 86.96 | − 37.95 | − 33.89 |
Ras signaling pathway | 9 | 39.13 | − 12.52 | − 8.76 |
PID TNF PATHWAY | 6 | 26.09 | − 11.87 | − 8.29 |
Regulation of kinase activity | 11 | 47.83 | − 11.26 | − 7.74 |
VEGFA-VEGFR2 signaling pathway | 9 | 39.13 | − 10.47 | − 7.11 |
Osteoclast differentiation | 6 | 26.09 | − 9.09 | − 6.05 |
Positive regulation of cell migration | 8 | 34.78 | − 8.08 | − 5.17 |
Focal adhesion | 6 | 26.09 | − 7.98 | − 5.12 |
ST P38 MAPK PATHWAY | 4 | 17.39 | − 7.66 | − 4.9 |
Signaling by receptor tyrosine kinases | 7 | 30.43 | − 6.87 | − 4.21 |
VEGF signaling pathway | 4 | 17.39 | − 6.83 | − 4.19 |
Platelet activation, signaling, and aggregation | 5 | 21.74 | − 5.7 | − 3.23 |
Actin filament-based process | 7 | 30.43 | − 5.63 | − 3.18 |
Cell junction organization | 6 | 26.09 | − 4.81 | − 2.5 |
Apoptosis modulation and signaling | 3 | 13.04 | − 4.23 | − 2.08 |
Hippo signaling regulation pathways | 3 | 13.04 | − 4.15 | − 2.02 |
Muscle structure development | 5 | 21.74 | − 3.86 | − 1.79 |
Negative regulation of secretion by cell | 3 | 13.04 | − 3.64 | − 1.61 |
Oxytocin signaling pathway | 3 | 13.04 | − 3.6 | − 1.57 |
Regulation of cell activation | 4 | 17.39 | − 2.76 | − 0.86 |