Expression data from Hematopoietic stem cells

Objective: Microarray analysis was used to determine the molecular mechanism underlying Fancd2 and Foxo3a double knockout mice HSCs exhaustion. Methods: SLAM cells were obtained from WT, Fancd2KO, Foxo3aKO and DKO bone marrow cells. Total RNA from SLAM cells was purified by RNeasy kit (Qiagen) following the manufacturer’s procedure. And then the RNA was fluorescently labeled and hybridized to Affmetrix Mogene 2.0 ST array. Data was analyzed by Genespring GX11 software. Results: a large proportion of differentially expressed genes in dKO HSCs belonged to cell cycle control. Another group of up-regulated genes were those involved in the differentiation of HSCs. Other differentially regulated transcripts included genes known to be involved in oxidative stress response, or to have roles in DNA repair. Conclusion: Cell cycle, DNA repair, DNA binding and hematopoietic lineage differentiation pathways might have been perturbed in HSC exhaustion. Mouse Femurs and tibias from WT, Fancd2KO, Foxo3aKO and DKO mice were flushed to dissociate the BM fraction. The mononuclear cells were isolated by Ficoll (GE Healthcare) gradient centrifugation. Single SLAM (Lin-ckit+Sca-1+CD34-) cells were sorted. RNA was extracted, labelled, and quantified using the Mouse ST-2 microarray.