Differentiation of mammary epithelial cells with/without exposure to RANKL.. Mus musculus

In order to investigate the mechanisms underlying defective alveologenesis caused by RANK over-expression in mice, global gene expression profiles from primary acinar cultures of mammary epithelial cells were analyzed. Overall design: Mammary epithelial cells from gestation day 16.5 WT and MMTV-RANK females were obtained. Cultures were established in the presence of prolactin, which induces differentiation into milk secreting acini, and with or without RANKL (Receptor Activator of NFkB Ligand) to precisely determine the role of RANK signaling at midgestation. Global gene expression profiling at 8 hours (h), 24 h and 72 h from WT, and MMTV-RANK 3D acinar cultures was obtained. Please note that experiments were performed technically as dual channel (e.g., Cy3 and Cy5-labeled samples hybridized to the same array; total 72 hybridizations) but the results were processed as though they were single channel. Rosetta Resolver ratio split algorithm was used to parse individual Cy3 (Ch1) or Cy5 (Ch2) intensity values and then to combine 4 replicate values for each sample into a single intensity value. Therefore, the normalized_combined_data.txt (available on the Series records) contains 36 data columns [i.e. (72array X 2 channel)/4 replicates; 2 genotype (WT or TG) X 3 replicates X 3 timepoints (8 ,24 ,72hr) X 2 treatments (w/&w/o RANKL)].