Identification of downstream targets of Ezh2 histone methyltransferase

Genome-wide transcriptional profiles of the mammary stem cell (MaSC)-enriched and luminal populations were determined following targeted disruption of Ezh2 gene. Gene ontology analysis revealed changes in the expression of genes associated with cell cycle, DNA replication and DNA repair. To carry out ex vivo cre-mediated excision of Ezh2, freshly isolated MaSC-enriched (CD29hiCD24+) and luminal (CD29loCD24+) cells from 8-week old Ezh2T/+ F/F mice were infected with virus expressing cre recombinase or control virus (for infection methods refer to Toula et. al., Cell stem cell 2008 11; 3(4):429-41). The infected cells were purified by sorting for GFP+ cells. Total RNA was isolated from GFP+ cells and up to 250 ng of total RNA per sample was labelled and hybridized to Illumina Mouse WG6V2 expression BeadChip platform. Three independent samples were used for each population.