Gene expression profile of mouse embryonic stem cells (D3-pOCT-mESC) grown in low concentrations of nitric oxide

In order to identify the genes regulated in mouse embryonic stem cells (mESC) by the effect of low concentrations of nitric oxide (NO), we analysed the transcriptome of cells treated with NO and compared it to those of cells cultured in the absence of leukemia inhibitory factor (LIF), and in the presence of LIF. We used the cell line D3-pOct4, which carries the enhanced Green Fluorescence Protein gene (eGFP) under the control of the Oct-4 promotor. This line is continuously maintained in the undifferentiated state in the presence of LIF, in comparison with the wild type line . D3-pOct4 cells cultured in the presence of 1000 U/ml of LIF, in the absence of LIF, and in the absence of LIF supplemented with 2uM of diethylenetriamine nitric oxide adduct (DETA)/NO (Sigma) used as donor of nitric oxide. Treatments were maintained for seven days. RNA was extracted and analyzed by Bioanalyzer of Agilent. The cDNA microarray chip used was Mouse Genome 430_2.0 (Affymetrix) and the data were analyzed with GeneChip Operating System v1.4.0.036 using global scaling as the normalization method.