Mesenchymal MACF1 Facilitates SMAD7 Nuclear Translocation to Drive Bone Formation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Gene Expression Profile
2.2. Human Bone Specimen
2.3. Generation of the MACF1 Conditional Knockout (cKO) Mouse
2.4. Study Design and Animal Grouping
2.5. Cell Preparation
2.6. RNA and Real Time Quantitative PCR (qPCR)
2.7. Protein and Western Blot
2.8. Immunofluorescence
2.9. Co-Immunoprecipitation and iTRAQ-LC-MS/MS
2.10. Cleared Skeleton Preparation
2.11. Bone Densitometry and Micro-Computed Tomography
2.12. Three-Point Bending Test
2.13. Histochemical, Calcein Labeling, and Histomorphometric Analysis
2.14. Serological Factors
2.15. Cell Culture Staining
2.16. Electric Cell-Substrate Impedance Sensing
2.17. Plasmid Preparation
2.18. Data Processing and Statistics
3. Results
3.1. Mesenchymal Microtubule Actin Crosslinking Factor 1 (MACF1) Is Decreased in Osteoporosis Patients
3.2. Mesenchymal Deletion of Microtubule Actin Crosslinking Factor 1 (MACF1)
3.3. Loss of Microtubule Actin Crosslinking Factor 1 (MACF1) Inhibits Osteogenic Differentiation in Mesenchymal Stem Cells (MSCs)
3.4. Mesenchymal Deletion of Microtubule Actin Crosslinking Factor 1 (MACF1) Impairs Early-Stage Bone Development in Mice
3.5. Microtubule Actin Crosslinking Factor 1 (MACF1) Is Required for Bone Formation
3.6. Loss of Microtubule Actin Crosslinking Factor 1 (MACF1) Weakens Bone Properties in Adult Mice
3.7. Microtubule Actin Crosslinking Factor 1 (MACF1) Interacts with SMAD Family Member 7 (SMAD7) in Mesenchymal Stem Cells (MSCs)
3.8. Microtubule Actin Crosslinking Factor 1 (MACF1) Facilitates Nucleus Translocation of SMAD Family Member 7 (SMAD7) to Drive Osteogenic Differentiation
4. Discussion
4.1. Microtubule Actin Crosslinking Factor 1 (MACF1) Is Essential for Regulating Bone Development and Formation
4.2. Microtubule Actin Crosslinking Factor 1 (MACF1) Interacts Directly with SMAD Family Member 7 (SMAD7) to Initiate Osteogenic Differentiation
4.3. Microtubule Actin Crosslinking Factor 1 (MACF1) Is a Novel Potential Target for Primary Osteoporosis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Zhao, F.; Ma, X.; Qiu, W.; Wang, P.; Zhang, R.; Chen, Z.; Su, P.; Zhang, Y.; Li, D.; Ma, J.; et al. Mesenchymal MACF1 Facilitates SMAD7 Nuclear Translocation to Drive Bone Formation. Cells 2020, 9, 616. https://doi.org/10.3390/cells9030616
Zhao F, Ma X, Qiu W, Wang P, Zhang R, Chen Z, Su P, Zhang Y, Li D, Ma J, et al. Mesenchymal MACF1 Facilitates SMAD7 Nuclear Translocation to Drive Bone Formation. Cells. 2020; 9(3):616. https://doi.org/10.3390/cells9030616
Chicago/Turabian StyleZhao, Fan, Xiaoli Ma, Wuxia Qiu, Pai Wang, Ru Zhang, Zhihao Chen, Peihong Su, Yan Zhang, Dijie Li, Jianhua Ma, and et al. 2020. "Mesenchymal MACF1 Facilitates SMAD7 Nuclear Translocation to Drive Bone Formation" Cells 9, no. 3: 616. https://doi.org/10.3390/cells9030616