Project description:Gene-level analysis of gene expression in immortalized mouse keratinocyte lines that express or lack integrin alpha3beta1 The laminin-332-binding integrin alpha3beta1 is expressed highly in the epidermis, but its roles in regulating gene expression programs that promote normal or pathological skin remodeling have not been underexplored previously. In order to identify genes that are regulated by alpha3beta1 in keratinocytes, we used microarray approaches to identify alpha3beta1-dependent genes in mouse keratinocytes that were immortalized with SV40 large T antigen (i.e., MK cells).
Project description:Gene-level analysis of gene expression in immortalized mouse keratinocyte lines that express integrins alpha3beta1 and alpha9beta1 in different combinations. The integrins alpha3beta1 and alpha9beta1 are expressed highly in the epidermis, but their roles in regulating gene expression programs that promote normal or pathological wound healing have not been explored previously. In order to identify genes that are regulated by these integrins in keratinocytes, we used microarray approaches to identify integrin-dependent genes in mouse keratinocytes that were immortalized with SV40 large T antigen (i.e., MK cells).
Project description:Nucleolus-associated DNA was isolated from MEF cells before and after conditional knock-out of UBF and hybridized against genomic DNA in biological replicates. Two different types of immortalized MEF cells were used. MEFs were immortalized by genetic depletion of p53, iMEFs were immortalized by transfection of the SV40 Tt antigen.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
