Loss of ZC4H2, an Arthrogryposis Multiplex Congenita Associated Gene, Promotes Osteoclastogenesis in Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal Experimentation
2.2. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
2.3. MSCs, BMDM Isolation and In Vitro Differentiation
2.4. RNA-Seq Analysis
2.5. Von Kossa Staining
2.6. Skeletal Staining
2.7. Micro-Computed Tomography (Micro-CT)
2.8. Alkaline Phosphatase (ALP) and Alizarin Red S (ARS) Staining
2.9. Tartrate Resistant Acid Phosphatase (TRAP) Staining Assay
2.10. F-Actin Ring Formation Assay
2.11. Scanning Electron Microscopy
2.12. ELISA Assay of CTX-1
2.13. Cell Counting Kit-8 (CCK-8) Assay
2.14. BrdU Incorporation Assay
2.15. Statistical Analysis
3. Results
3.1. Loss of ZC4H2 Reduces Long Bone Calcification in Mice
3.2. Loss of ZC4H2 Reduces Femoral Bone Mass
3.3. Loss of ZC4H2 Does Not Affect Osteoblast Differentiation
3.4. Loss of ZC4H2 Promotes Osteoclastogenesis and Bone Resorption
3.5. Loss of ZC4H2 Inhibits Proliferation of MSCs and BMDM
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Forward Sequence | Reverse Sequence |
---|---|---|
ACTB | GGCTGTATTCCCCTCCATCG | CCAGTTGGTAACAATGCCATGT |
ZC4H2 | AAAGATCAAGGCCCGTTTG | TTGTATTCTTTCAGGTGCCTCTC |
OSX | ACTCATCCCTATGGCTCGTG | GGTAGGGAGCTGGGTTAAGG |
OCN | ACTCCGGCGCTACCTTGGAGCC | GCAGGGTTAAGCTCACACTG |
Runx2 | CCTAGTTAGAGTGGTAGCAGA | ACAGACAACGAAGAAAGTTCC |
Col1a1 | GCTCCTCTTAGGGGCCACT | CCACGTCTCACCATTGGGG |
Ctsk | CTCGGCGTTTAATTTGGGAGA | TCGAGAGGGAGGTATTCTGAGT |
Mmp9 | CTGGACAGCCAGACACTAAAG | CTCGCGGCAAGTCTTCAGAG |
Acp5 | CACTCCCACCCTGAGATTTGT | CATCGTCTGCACGGTTCTG |
Dcstamp | GGGGACTTATGTGTTTCCACG | ACAAAGCAACAGACTCCCAAAT |
Ocstamp | CTGTAACGAACTACTGACCCAG | CCCAGGCTTAGGAAGACGAAG |
Atp6v0d2 | CAGAGCTGTACTTCAATGTGGA | AGGTCTCACACTGCACTAGGT |
Oscar | CCTAGCCTCATACCCCCAG | CGTTGATCCCAGGAGTCACAA |
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Zhu, L.; Zhang, L.; Cha, J.; Li, C.; Mao, B. Loss of ZC4H2, an Arthrogryposis Multiplex Congenita Associated Gene, Promotes Osteoclastogenesis in Mice. Genes 2024, 15, 1134. https://doi.org/10.3390/genes15091134
Zhu L, Zhang L, Cha J, Li C, Mao B. Loss of ZC4H2, an Arthrogryposis Multiplex Congenita Associated Gene, Promotes Osteoclastogenesis in Mice. Genes. 2024; 15(9):1134. https://doi.org/10.3390/genes15091134
Chicago/Turabian StyleZhu, Liang, Longlong Zhang, Jingmei Cha, Chaocui Li, and Bingyu Mao. 2024. "Loss of ZC4H2, an Arthrogryposis Multiplex Congenita Associated Gene, Promotes Osteoclastogenesis in Mice" Genes 15, no. 9: 1134. https://doi.org/10.3390/genes15091134