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Volumn 15, Issue 17, 2005, Pages 3948-3952

Induction of apoptosis by a novel indirubin-5-nitro-3′-monoxime, a CDK inhibitor, in human lung cancer cells

Author keywords

A549 human lung cancer cells; Apoptosis; Indirubin 5 nitro 3 monoxime; p21; p53

Indexed keywords

CYCLIN DEPENDENT KINASE INHIBITOR; INDIRUBIN; INDIRUBIN 3' MONOXIME; INDIRUBIN 5 NITRO 3' MONOXIME; INDIRUBIN 5 SULFONATE; PROTEIN P53; UNCLASSIFIED DRUG;

EID: 23244449448     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2005.05.105     Document Type: Article
Times cited : (52)

References (33)
  • 21
    • 33544462775 scopus 로고    scopus 로고
    • note
    • 6 cells in a 100 mm dish) were seeded for 24 h and then incubated with test samples for additional 24 h. Floating and attached cells were combined, washed with ice-cold PBS, and then fixed in a 4% paraformaldehyde solution at 4°C for 10 min. Fixed cells were placed on slides and stained with Hoechst 33258 (0.3 μg/ml) at room temperature for 10 min in the dark. Stained cells were mounted with 80% glycerol, and then visualized using fluorescence microscope.
  • 22
    • 33544455669 scopus 로고    scopus 로고
    • note
    • DNA fragmentation: A549 cells were treated with test samples for 24 h. After incubation, floating and attached cells were combined by centrifugation and trypsinization. The combined cells were washed with ice-cold PBS and lysed in 100 μl of DNA isolation buffer [50 mM Tris-HCl (pH 7.5), 20 mM EDTA, 1% NP-40]. After centrifugation, 10 μl of 10% SDS solution (final 1%) and 10 μl of 50 mg/ml RNase A (final 5 μg/ml) were added to the supernatants, and incubated at 56°C for 1 h. Subsequently, proteinase K (5 μg/ml) was added and then incubated at 37°C for 2 h. DNA was precipitated with 0.5 volume of 10 M ammonium acetate and 2.5 volume of cold ethanol at -70°C overnight. Precipitated DNA was dissolved in 50 μl of Tris-EDTA buffer (pH 8.0). The same concentration of DNA samples (5 μg) were resolved by electrophoresis in 2% agarose gel at 60 V for 2 h, stained with SYBR Gold (Molecular Probes, Eugene, OR, USA) for 1 h in the dark, and visualized under the UV transilluminator.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.