Gene expression in mouse mammary cells: whole mammary gland, EpCAM-Int and CD29-High subset and lung-colonizing cells. Mus musculus

The experiment was carried out to identify gene expression changes between mammary cells that colonize lungs following intravenous delivery, a subset of the dissociated mammary cells that contains lung colonizing subset, and the intact mammary gland. Overall design: The experiment was designed to determine LCC gene expression profile, and to identify differentially expessed genes between LCC, E-Int29-Hi, and MG samples. For each experiment, mammary glands from 8 FVB.GFP females were dissociated and injected i.v. in two 2.5x10^6 doses into 4 FVB recipients. One day after injection, the lungs of recipient mice were dissociated and GFP or DDAO-stained cells isolated by a MoFlo (Beckman Coulter) flow sorter. The RNA from pooled LCC sample and the controls was extracted using MagMAXtm-96 Total RNA Isolation kit (AM1830, Ambion) according to manufacturer’s instructions. Best quality RNA as determined by 18S/28S fluorescence (Agilent 2100) was used for the sequencing experiment, and other preparations were used for RT-PCR applications.