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Blood, Vol. 107, Issue 3, 1174-1177, February 1, 2006
The novel DNA methylation inhibitor zebularine is effective against the development of murine T-cell lymphoma
Blood Herranz et al.
107: 1174
Supplemental materials for: Herranz et al, Vol 107, Issue 3, 1174-1177
Files in this Data Supplement:
- Figure S1. Chromosomal analysis and zebularine use (JPG, 485 KB)
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The top panel shows the spectral karyotype analysis (SKY) of lymphoid cells from a zebularine-treated mouse. The image shows a representative cell with 40 chromosomes and an absence of structural aberrations. The bottom panel shows G-banded chromosomes from the same mouse.
- Figure S2. Bisulfite genomic sequencing analysis for p16INK4a, MLT-1, E-cadherin and MGMT CpG island promoter methylation and zebularine treatment (JPG, 754 KB)
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The left-hand columns provide representative electropherograms of a clone of bisulfite-modified DNA from the corresponding promoter-containing CpG island regions. The presence of protected Cs indicates methylated CpG sites. The center columns show schematic representations of the methylation status of each CpG dinucleotide in the bisulfite genomic sequencing of 10 clones. Black and white dots indicate methylated and unmethylated CpGs, respectively. The black arrows indicate the position of the transcription start sites. The right-hand columns show p16INK4a and MLT-1 expression analysis by reverse-transcription PCR.
- Figure S3. The DNA demethylating effects of zebularine in a MOLT-4 xenograft (JPG, 340 KB)
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(A) High-performance capillary electropheresis analysis, showing a reduction of 5-methylcytosine genomic content in the MOLT-4 zebularine-treated xenograft. (B) Schematic bisulfite genomic sequencing analysis for RASSF1A and p15INK4b CpG island promoter methylation before and after zebularine treatment. Four clones are represented. Black and white dots indicate methylated and unmethylated CpGs, respectively. The black arrows indicate the position of the transcription start sites. (C) Western blot showing depletion of DNMT1 protein levels after zebularine treatment in the native (MOLT-4 ZEB) and xenografted (MOLT-4 X-ZEB) MOLT-4 cell lines.
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