Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover
Abstract
1. Introduction
2. Results
2.1. Cross-Species Analyses and Single-cell Quantitative Imaging Microscopy Identify Putative Human CIN Genes
2.2. Putative CIN Genes Exhibit Frequent Copy Number Losses that may Be Pathogenic in Cancer
2.3. Reduced Gene Expression Drives Increases in Chromosome Aberrations
2.4. Reduced SKP1 Expression Induces Replication Stress and DNA Double-strand Breaks
2.5. SKP1 Silencing Underlies Aberrant Increases in the Oncoprotein Cyclin E1
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Culture
4.2. Identification of Candidate Human CIN Genes
4.3. Gene Silencing
4.4. ScQuantIM
4.5. Gene Alterations and Outcome Analyses in Cancer
4.6. Mitotic Chromosome Spreads and Chromosome Enumeration
4.7. Indirect Immunofluorescent Labeling and Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Target Protein 1 | Species | Catalog No. | [WB] 2 | [IIF] 3 |
---|---|---|---|---|
α-Tubulin * | Mouse | Abcam, ab7291 | 1:4000 | - |
ARL2 | Rabbit | Abcam; ab183510 | 1:5000 | - |
BUB3 | Rabbit | Abcam; ab4180 | 1:5000 | - |
CCNE1 | Rabbit | Abcam; ab133266 | 1:5000 | 1:200 |
Cyclophilin B * | Rabbit | Abcam; ab16045 | 1:50,000 | - |
DSN1 | Rabbit | Thermo Fisher; PA534879 | 1:1000 | - |
GARS | Rabbit | Abcam; ab42905 | 1:5000 | - |
GART | Rabbit | Abcam; ab169550 | 1:5000 | - |
γH2AX | Rabbit | Abcam; ab2893 | - | 1:1000 |
NUF2 | Rabbit | Abcam; ab176556 | 1:20,000 | - |
PIGS | Rabbit | Abcam; ab113817 | 1:5000 | - |
RPA32/RPA2 | Mouse | Abcam; ab2175 | - | 1:200 |
SHMT2 | Rabbit | Abcam; ab180786 | 1:1000 | - |
SKP1 | Mouse | Abcam; ab124473 | 1:5000 | - |
SPC24 | Rabbit | Abcam; ab157184 | 1:5000 | - |
Anti-Rabbit HRP | Goat | Jackson Immunoresearch; 111-035-114 | 1:10,000 | - |
Anti-Mouse HRP | Goat | Jackson Immunoresearch; 115-035-146 | 1:10,000 | - |
Anti-Rabbit AlexFluor488 | Goat | Abcam; ab150081 | - | 1:200 |
Anti-Mouse Cy3 | Goat | Abcam; ab150117 | - | 1:200 |
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Thompson, L.L.; Baergen, A.K.; Lichtensztejn, Z.; McManus, K.J. Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover. Cancers 2020, 12, 531. https://doi.org/10.3390/cancers12030531
Thompson LL, Baergen AK, Lichtensztejn Z, McManus KJ. Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover. Cancers. 2020; 12(3):531. https://doi.org/10.3390/cancers12030531
Chicago/Turabian StyleThompson, Laura L., Allison K. Baergen, Zelda Lichtensztejn, and Kirk J. McManus. 2020. "Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover" Cancers 12, no. 3: 531. https://doi.org/10.3390/cancers12030531
APA StyleThompson, L. L., Baergen, A. K., Lichtensztejn, Z., & McManus, K. J. (2020). Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover. Cancers, 12(3), 531. https://doi.org/10.3390/cancers12030531