Loss of Dystroglycan Drives Cellular Senescence via Defective Mitosis-Mediated Genomic Instability
Abstract
:1. Introduction
2. Results
2.1. Generation and Characterization of CRISPR/Cas9-Mediated DG-Null C2C12 Cell Clones
2.2. DG Deficiency Provokes Altered Localization and Decreased Protein Levels of Lamin B1
2.3. The Loss of DG Induces the Expression of Senescence-Associated Features
2.4. Aberrant Multipolar Mitoses in DG-KO Cells Resulted in Micronuclei Formation and Activation of a P53-Mediated DNA Damage Response
3. Discussion
4. Materials and Methods
4.1. Cell Culturing and Treatments
4.2. Generation of DG-KO C2C12 Cell Lines by CRISPR-Cas
4.3. Antibodies
4.4. Western Blotting
4.5. Immunofluorescence and Confocal Microscopy Analysis
4.6. Flow Cytometry and Cell Proliferation Assays
4.7. Fluorescence in Situ Hybridization (FISH) and Relative Telomere Length Determination
4.8. Senescence-Associated β-Galactosidase (SA-β-Gal) Assay
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
NaBu | Sodium butyrate |
PM | Plasma membrane |
NE | Nuclear envelope |
DAPs | Dystrophin-Associated Proteins |
ECM | Extracellular matrix |
SA-β-gal | Senescence associated β-galactosidase |
FACs | Fluorescence-activated cell sorting |
SDS PAGE | Sodium dodecyl sulfate polyacrylamide gel |
CLSM | Confocal laser scanning microscopy analysis |
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Jimenez-Gutierrez, G.E.; Mondragon-Gonzalez, R.; Soto-Ponce, L.A.; Gómez-Monsiváis, W.L.; García-Aguirre, I.; Pacheco-Rivera, R.A.; Suárez-Sánchez, R.; Brancaccio, A.; Magaña, J.J.; C.R. Perlingeiro, R.; et al. Loss of Dystroglycan Drives Cellular Senescence via Defective Mitosis-Mediated Genomic Instability. Int. J. Mol. Sci. 2020, 21, 4961. https://doi.org/10.3390/ijms21144961
Jimenez-Gutierrez GE, Mondragon-Gonzalez R, Soto-Ponce LA, Gómez-Monsiváis WL, García-Aguirre I, Pacheco-Rivera RA, Suárez-Sánchez R, Brancaccio A, Magaña JJ, C.R. Perlingeiro R, et al. Loss of Dystroglycan Drives Cellular Senescence via Defective Mitosis-Mediated Genomic Instability. International Journal of Molecular Sciences. 2020; 21(14):4961. https://doi.org/10.3390/ijms21144961
Chicago/Turabian StyleJimenez-Gutierrez, Guadalupe Elizabeth, Ricardo Mondragon-Gonzalez, Luz Adriana Soto-Ponce, Wendy Lilián Gómez-Monsiváis, Ian García-Aguirre, Ruth Abigail Pacheco-Rivera, Rocío Suárez-Sánchez, Andrea Brancaccio, Jonathan Javier Magaña, Rita C.R. Perlingeiro, and et al. 2020. "Loss of Dystroglycan Drives Cellular Senescence via Defective Mitosis-Mediated Genomic Instability" International Journal of Molecular Sciences 21, no. 14: 4961. https://doi.org/10.3390/ijms21144961