Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells
Abstract
:1. Introduction
2. Results
2.1. The Kinetics of Focus Formation and Decay
2.2. Focus Area
2.3. Focus Intensity Dynamics
2.4. Focus Mobility
2.5. Focus Merging and Splitting
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Cell Line Authentication
4.2. Irradiation
4.3. Live Cell Imaging
4.4. Image Analysis
4.5. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Intrest
Abbreviations
DSB | DNA double strand breaks |
FPC | frequency per cell |
GFP | green fluorescence protein |
HRR | homologous recombination repair |
LET | linear energy transfer |
NHEJ | non-homologous end joining |
p.r. | post radiationem |
SD | standard deviation |
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Radiation Type | Initial FPC | Highest FPC (@ Time Point) | Final FPC | Max. Focus Increase 1 | Max. Focus Reduction 2 |
---|---|---|---|---|---|
Control | 1.5 ± 1.5 | 1.5 ± 1.5 | 1.5 ± 1.5 | - | - |
Alpha particles | 6.4 ± 3.1 | 8.1 ± 2.2 (11 min) | 6.9 ± 3.1 | 5.4 | 1.2 |
X-rays | 9.7 ± 4.3 | 15.3 ± 5.8 * (16 min) | 8.7 ± 4.2 | 10.2 | 1.8 |
Mixed beams | 7.6 ± 3.4 | 10.3 ± 4.8 (9 min) | 9.1 ± 2.8 | 6.8 | 1.1 |
Expected | 8.05 | 11.7 (15 min) | 7.8 | 7.8 | 1.3 |
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Sollazzo, A.; Brzozowska, B.; Cheng, L.; Lundholm, L.; Scherthan, H.; Wojcik, A. Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells. Int. J. Mol. Sci. 2018, 19, 519. https://doi.org/10.3390/ijms19020519
Sollazzo A, Brzozowska B, Cheng L, Lundholm L, Scherthan H, Wojcik A. Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells. International Journal of Molecular Sciences. 2018; 19(2):519. https://doi.org/10.3390/ijms19020519
Chicago/Turabian StyleSollazzo, Alice, Beata Brzozowska, Lei Cheng, Lovisa Lundholm, Harry Scherthan, and Andrzej Wojcik. 2018. "Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells" International Journal of Molecular Sciences 19, no. 2: 519. https://doi.org/10.3390/ijms19020519