Project description:c-jun conditional knockout mice (c-jun?li) and relative controls (c-junf/f) were treated with TCPOBOP (3mg/Kg of body weight) and were sacrificed at 0, 1, 3 and 6 hours later. Gene expression analysis was performed on total RNA pools extracted from fresh frozen livers of 3 animals per group. For each time point 4 technical replicates were analyzed except for the 6 hours samples which were in triplicate.
Project description:Constitutive signalling pathway activation is a key feature of primary tumours and cancer cell lines. The regulation of gene expression changes may be via both genomic and epigenetic means, and understanding the mechanisms by which signal transduction may be activated can provide rational targets for therapeutic intervention. We have previously carried out DNA copy and expression profiling on a unique panel of five paediatric glioma cell lines, and have noted only a limited influence of gene amplification on gene expression. In the present study we have extended our work to include a measure of global methylation changes as well as micro RNA profiling in the same cell lines. We noted various instances of signalling pathway activation associated with specific hypermethylation or miRNA regulation of gene expression at various effectors also observed in primary paediatric tumours. These data provide evidence for the multifaceted nature of gene expression changes in paediatric high grade glioma, and identify novel targets for targeted therapy in this treatment refractory disease.
Project description:To evaluate the experimental performance of Actichip microarrays, a series of 10 repeated hybridisation experiments, including 5 dye swaps, were carried out with optimised target labelling, hybridisation, scanning and data analysis protocols. All the procedures were standardised to limit the impact of experimental bias or biological variations on data. The same series of high quality RNA samples purified from human breast adenocarcinoma MCF-7 cells and from human skeletal muscle was used in our experiments. Epithelial cells and muscle tissue were chosen because they express well-characterised sets of cytoskeleton genes, and were anticipated to give well-contrasted differential expression data when analysed with Actichip.
Project description:Gene expression was measured in control and heat resistance selected adult female flies before and at 8 time points after heat stress for 1h @ 36 degrees,Abstract The availability of full genome sequences has allowed the construction of microarrays, with which screening,of the full genome for changes in gene expression is possible. This method can provide a wealth of information about,biology at the level of gene expression and is a powerful method to identify genes and pathways involved in various,processes. In this study, we report a detailed analysis of the full heat stress response in Drosophila melanogaster,females, using whole genome gene expression arrays (Affymetrix Inc, Santa Clara, CA, USA). The study focuses on,up- as well as downregulation of genes from just before and at 8 time points after an application of short heat hardening,(368C for 1 hour). The expression changes were followed up to 64 hours after the heat stress, using 4 biological,replicates. This study describes in detail the dramatic change in gene expression over time induced by a short-term,heat treatment. We found both known stress responding genes and new candidate genes, and processes to be involved,in the stress response. We identified 3 main groups of stress responsive genes that were early–upregulated, early–,downregulated, and late–upregulated, respectively, among 1222 differentially expressed genes in the data set. Comparisons,with stress sensitive genes identified by studies of responses to other types of stress allow the discussion of,heat-specific and general stress responses in Drosophila. Several unexpected features were revealed by this analysis,,which suggests that novel pathways and mechanisms are involved in the responses to heat stress and to stress in,general. The majority of stress responsive genes identified in this and other studies were downregulated, and the,degree of overlap among downregulated genes was relatively high, whereas genes responding by upregulation to heat,and other stress factors were more specific to the stress applied or to the conditions of the particular study. As an,expected exception, heat shock genes were generally found to be upregulated by stress in general.
Project description:rs06-06_mirna - flg22 treatment & mutants 2 - Time course : What are the genes (incuding microRNA precursors) that are differentially regulated upon flg22 treatment? miRNA : What are the genes (including miRNA precursors) that are differentially regulated in a set of microRNA mutants? siRNA : What are the genes (including miRNA precursors) that are differentially regulated in a set of siRNA mutants? - Col-0, dcl4-1, rdr6-1 were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium 2 days after and treated with 10uM flagellin active or inactive. Samples were harvested at 30 min after treatment. Keywords: treated vs untreated comparison 3 dye-swap - CATMA arrays