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Volumn 287, Issue 5462, 2000, Pages 2486-2492

Mitotic misregulation and human aging

Author keywords

[No Author keywords available]

Indexed keywords

MESSENGER RNA;

EID: 0034737456     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.287.5462.2486     Document Type: Article
Times cited : (512)

References (49)
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    • 2 at 37°C and were split in a 1:2 ratio at confluency with trypsin-EDTA. Bromodeoxyuridine (BrdU) incorporation was performed according to Becton Dickinson (Franklin Lakes, NJ) protocols and imaged with epifluorescence microscopy. After a 48-hour incubation with 10 μM BrdU and staining with antibody to BrdU-fluorescein isothiocyanate (FITC), more than 90% of the fibroblasts derived from old-age and progeria individuals showed intense staining of the nucleus, a similar ratio compared with cells from young and middle-age individuals. The analysis was based on 200 cells counted.
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    • note
    • Cells were grown under identical conditions for three additional doublings to about 60% confluency. Polyadenylated RNAs were isolated and converted to cDNA, followed by in vitro transcription cRNA synthesis. The biotinylated cRNAs were then hybridized to the HuGeneFL oligonucleotide arrays (Affymetrix, Santa Clara, CA), which contain probes for more than 6000 known human genes. All procedures were performed according to prescribed Affymetrix protocols.
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    • note
    • We thank the Skaggs Institute for Chemical Sciences and the American Chemical Society Irving Sigal Post-doctoral Fellowship (D.L.) for financial support, D. Wemmer and G. Rosania for helpful discussions, A. Su for computational support, K. Sullivan for assistance with microscopy, and J. Trotter for help with flow cytometry.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.