Alternative Splicing of Pericentrin Contributes to Cell Cycle Control in Cardiomyocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation and Cell Culture of Postnatal Cardiomyocytes
2.2. Cell Culture of C2C12 Myoblasts and ARPE-19 Adult Retinal Pigment Epithelial Cells
2.3. SiRNA Knockdown
2.4. Analysis of DNA Incorporation
2.5. Reverse Transcriptase PCR (RT-PCR)
2.6. Plasmids
2.7. Plasmid Transfection
2.8. Immunofluorescence and Microscopy
2.9. Image Preparation and Analysis
2.10. Statistical Analysis
3. Results
3.1. Pcnt B Preferentially Localizes at the Centrosome and Pcnt S at the Nuclear Envelope
3.2. Pcnt S Is Not Required for Centriole Cohesion
3.3. Ectopic Expression of Pcnt S Induces Centriole Splitting and Centriolar γ-Tubulin Reduction
3.4. Ectopic Pcnt S Expression Impairs DNA Synthesis
3.5. Pcnt S Depletion Enhances Serum-Induced Cell Cycle Re-Entry in Cardiomyocytes
3.6. Pcnt S Depletion Enhances Serum-Induced Cell Division in Cardiomyocytes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Steinfeldt, J.; Becker, R.; Vergarajauregui, S.; Engel, F.B. Alternative Splicing of Pericentrin Contributes to Cell Cycle Control in Cardiomyocytes. J. Cardiovasc. Dev. Dis. 2021, 8, 87. https://doi.org/10.3390/jcdd8080087
Steinfeldt J, Becker R, Vergarajauregui S, Engel FB. Alternative Splicing of Pericentrin Contributes to Cell Cycle Control in Cardiomyocytes. Journal of Cardiovascular Development and Disease. 2021; 8(8):87. https://doi.org/10.3390/jcdd8080087
Chicago/Turabian StyleSteinfeldt, Jakob, Robert Becker, Silvia Vergarajauregui, and Felix B. Engel. 2021. "Alternative Splicing of Pericentrin Contributes to Cell Cycle Control in Cardiomyocytes" Journal of Cardiovascular Development and Disease 8, no. 8: 87. https://doi.org/10.3390/jcdd8080087