Transcriptome profiling reveals novel Disabled-2-associated downstream effectors in mouse embryonic stem cells (ESC). Mus musculus

Disabled-2 (DAB2) is up-regulated when ESC were cocultured with OP9 stromal cells for mesodermal colony formation. Expression of DAB2 short hairpin small interfering RNA (shDAB2) altered ESC cell-cell adhesion that subsequently affected colony formation and mesoderm differentiation competence. To further unveil the molecular mechanisms associated with the phenotypic change of shDAB2-expressing ES cells, comprehensive profiling of DAB2-regulated molecules in shDAB2-ES cells (ES-V9 and 285-4) were performed by Affimatrix microarray. Overall design: Total RNAs from vector control (ES-V9) and shDAB2-expressing (285-4) ESC were isolated by Trizol (Invitrogen) and were further purified using the RNeasy mini kit (Qiagen). For profiling of mRNA expression, the purified total RNAs were then subjected to microarray analysis (Affymetrix gene chip mouse 430A).