Dataset features


Application: Gene expression microarray analysis, DNA methylation array analysis
Number of samples: 4
Release date: Apr 5 2007
Last update date: Oct 29 2018
Access: Public
Diseases: Neoplasms, Osteosarcoma
Dataset link Modulation of gene expression by decitabine in U-2OS cells in vitro and in vivo

Experimental Protocol

Total RNA was extracted using the RNeasy kit (Qiagen, Germany) from duplicate experiments of U-2OS cells at Day3 after treatment with 1 µM decitabine or medium alone (control). In each experiment RNA yields were pooled from two independent cultures per treatment arm to minimize experimental noise. For each case, 10 µg of RNA was labeled and hybridized to the Affymetrix HG-U133A GeneChips using the manufacturer’s protocol (Affymetrix, Santa Clara, CA) by the Center of Applied Genomics at the Hospital for Sick Children (Toronto, Canada). Data were extracted using the Microarray Suite (MAS) version 5.0 (Affymetrix) and linearly scaled to achieve an average intensity of 150 across each chip. The candidate gene list obtained from the MAS 5.0-extracted data was selected by eliminating genes that were not present in at least one experiment. The arrays were subjected to a pair wise comparison using MAS 5.0, with signal intensities from the no-treatment cells as the baseline. The statistical significance for the change of expression for each probe set between the decitabine treated and control was calculated by the MAS5.0 software. The criteria for gene selection for real-time expression validation analysis was based on the statistically significant up-regulation (p<0.0025) and fold change of ≥ 2 for expression after decitabine treatment. The gene list was annotated based on the NetAffx data-base ( /index.affx) and further verified using the Human Genome Browser data base (










Khaldoun Al-Romaih

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