Twist1 in skin tumorigenesis

This study was designed to investigate the transcripts that are regulated by Twist1 in skin tymor epithelial cells in a p53-dependent and independent manner. To this aim, Tumor epithelial cells from primary mouse skin tumors of different genotypes were FACS sorted and analyzed by microarray. Tumor bearing mice of different genotypes (CTRL, K14CreER:Twist1fl/fl, K14CreER:Twist1fl/fl:p53fl/fl) were treated with tamoxifen for 5 days to induce the deletion of Twist1 with or without p53 and. Tumors were disected and digested to harvest tumor epithelial cells. Tumor cells were FACS sorted using a combination of Epcam/CD45/CD31/CD140a and DAPI ti isolate tumor epithelial cells. These cells were harvested in lysis buffer and RNA was extracted using the Qiagen RNA purification microkit to perform microarrayprofilling. All the mice used for this study are all littermates from mixed background strains (up to 6 transgenes combined). The details relative to these mice are described in Cell Stem Cell article (in press).