Zonula occludens 2 and Cell-Cell Contacts Are Required for Normal Nuclear Shape in Epithelia
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Quantification of Nuclear Circularity
2.3. Transmission Electron Microscopy
2.4. DNA Transfection
2.5. Immunofluorescence
2.6. Nuclear Fluorescence Quantification with ImageJ
2.7. Western Blots
2.8. Western Blot Densitometry with ImageJ
2.9. Immunofluorescence Detection of β-Galactosidase Activity
2.10. Immunofluorescence Detection of Apoptosis
2.11. Pull-Down Assays
2.12. Cell Fractionation Assay
2.13. Drugs
2.14. Statistical Analysis
3. Results
3.1. In ZO-2 KD MDCK Cells, the Nuclei Are Lobulated and Display Multiple Indentations
3.2. The Lack of ZO-2 Increases the Phosphorylation of Histone H2AX, Indicative of More Abundant DNA Double Strands Breaks
3.3. ZO-2 KD Cells Are More Resistant to Senescence Than Parental Cells
3.4. In ZO-2 KD MDCK Cells, the Expression of Lamins A/C and B, and Lamin B Receptor Is Unaffected
3.5. The Lack of ZO-2 Induces a Decrease in Constitutive and Facultative Heterochromatin in MDCK Cells
3.6. The Nuclei of ZO-2 KD MDCK Cells Are Lobulated and Present Indentations Due to the Instability of Microtubules
3.7. ZO-2 Is Needed for the Establishment of the Microtubule Network between the Nucleus and the TJs That Maintains the Nuclear Shape
3.8. ZO-2 Associates to SUN-1 and Lamin B1, and Its Absence Alters the Expression of Nesprins-3 and -4 and Vimentin
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Hernández-Guzmán, C.; Gallego-Gutiérrez, H.; Chávez-Munguía, B.; Martín-Tapia, D.; González-Mariscal, L. Zonula occludens 2 and Cell-Cell Contacts Are Required for Normal Nuclear Shape in Epithelia. Cells 2021, 10, 2568. https://doi.org/10.3390/cells10102568
Hernández-Guzmán C, Gallego-Gutiérrez H, Chávez-Munguía B, Martín-Tapia D, González-Mariscal L. Zonula occludens 2 and Cell-Cell Contacts Are Required for Normal Nuclear Shape in Epithelia. Cells. 2021; 10(10):2568. https://doi.org/10.3390/cells10102568
Chicago/Turabian StyleHernández-Guzmán, Christian, Helios Gallego-Gutiérrez, Bibiana Chávez-Munguía, Dolores Martín-Tapia, and Lorenza González-Mariscal. 2021. "Zonula occludens 2 and Cell-Cell Contacts Are Required for Normal Nuclear Shape in Epithelia" Cells 10, no. 10: 2568. https://doi.org/10.3390/cells10102568