Project description:In order to establish a profile of gene expression of tumours treated with sunitinib, 4T1 breast tumours were allowed to establish in Balb/C mice for 1 week. The mice were then treated with 40 mg/kg Sunitinib in PBS and 3.72% DMSO (treatemt group) or PBS-DMSO vehicle alone (no treatment group), daily for the period of the experiment, after which the tumours were resected homogenised and subjected to CD31 microbead separation of endothelial cells and bulk tissue, both of which were subjected to agilent microarray analysis. Treated tumours greater than 250 mm3 (by caliper measurement) after 8 days of treatment were considered resistant to treatment, those smaller than this were considered sensitive to sunitinib treatment.
Project description:In order to establish a profile of gene expression of tumours treated with sunitinib, 4T1 breast tumours were allowed to establish in Balb/C mice for 1 week. The mice were then treated with 40 mg/kg Sunitinib in PBS and 3.72% DMSO (treatemt group) or PBS-DMSO vehicle alone (no treatment group), daily for the period of the experiment, after which the tumours were resected homogenised and subjected to CD31 microbead separation of endothelial cells and bulk tissue, both of which were subjected to agilent microarray analysis. Treated tumours greater than 250 mm3 (by caliper measurement) after 8 days of treatment were considered resistant to treatment, those smaller than this were considered sensitive to sunitinib treatment. Total RNA was extracted from each tumour sample using the Qiagen miRNAeasy kit, processed into cRNA using the Agilient low input, one colour labelling kit, hybridised onto the 8 x 66k whole mouse genome expression microarrays. Data was normalised in R64 and comparison matrices between treated and untreated endothelum and bulk fractions were constructed. Genes whose expression was significantly altered by sunitinib treatement were taken forward for further analysis.
Project description:The importance of unanchored Ub in innate immunity has been shown only for a limited number of unanchored Ub-interactors. We investigated what additional cellular factors interact with unanchored Ub and whether unanchored Ub plays a broader role in innate immunity. To identify unanchored Ub-interacting factors from murine lungs, we used His-tagged recombinant poly-Ub chains as bait. These chains were mixed with lung tissue lysates and protein complexes were isolated with Ni-NTA beads. Sample elutions were subjected to mass spectrometry (LC-MSMS) analysis.
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