NEIL1 and NEIL2 Are Recruited as Potential Backup for OGG1 upon OGG1 Depletion or Inhibition by TH5487
Abstract
:1. Introduction
2. Results
2.1. OGG1 Inhibitor TH5487 Does Not Bind to NEIL1
2.2. TH5487 Treatment Results in Increased Recruitment of NEIL1-GFP to DNA Damage Sites
2.3. NEIL1-Chromatin Binding Increases upon DNA Damage Induction and TH5487 Treatment
2.4. NEIL1 Retention Is Increased in a Dose-Dependent Manner upon DNA Damage Induction and TH5487 Treatment
2.5. TH5487 Treatment Results in Prolonged Accumualtion of NEIL2-GFP at DNA Damage Sites
2.6. NEIL1 and NEIL2 Nuclear Retention Increases upon DNA Damage Induction in OGG1-Depleted Cells
2.7. TH5487 Leads to 8-oxoG Lesion Accumulation upon Induction of Oxidative Stress in NEIL1-or NEIL2-Depleted Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatments
4.2. Differential Scanning Fluorimetry (DSF)
4.3. NEIL1-GFP and NEIL2-GFP Plasmids Construction
4.4. Live Cell Microscopy, Laser Microirradiation and Fluorescence Recovery after Photobleaching
4.5. Quantitative Microscopy
4.6. OGG1 Knockdown and In Situ Extraction
4.7. Western Blot
4.8. NEIL1 and NEIL2 Knockdown
4.9. Cell Viability Assay
4.10. Real-Time PCR
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Hanna, B.M.F.; Michel, M.; Helleday, T.; Mortusewicz, O. NEIL1 and NEIL2 Are Recruited as Potential Backup for OGG1 upon OGG1 Depletion or Inhibition by TH5487. Int. J. Mol. Sci. 2021, 22, 4542. https://doi.org/10.3390/ijms22094542
Hanna BMF, Michel M, Helleday T, Mortusewicz O. NEIL1 and NEIL2 Are Recruited as Potential Backup for OGG1 upon OGG1 Depletion or Inhibition by TH5487. International Journal of Molecular Sciences. 2021; 22(9):4542. https://doi.org/10.3390/ijms22094542
Chicago/Turabian StyleHanna, Bishoy M. F., Maurice Michel, Thomas Helleday, and Oliver Mortusewicz. 2021. "NEIL1 and NEIL2 Are Recruited as Potential Backup for OGG1 upon OGG1 Depletion or Inhibition by TH5487" International Journal of Molecular Sciences 22, no. 9: 4542. https://doi.org/10.3390/ijms22094542