Project description:We have employed chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) to analyze changes in chromatin architecture as well as the occupancy of two RNA Polymerase II (RNAPII) isoforms, initiation-competent (RNAPIIS5ph) as well as elongation-competent (RNAPIIS2ph) upon stress induction. We used resting mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway.
Project description:We have employed short-capped RNA sequencing (sc-RNA-seq) in order to identify genes whose expression is regulated by promoter proximal pausing of RNA Polymerase II (RNAPI) in response to stress stimulation. We used serum-deprived mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway.
Project description:Contemporary high dimensional biological assays, such as mRNA expression microarrays, regularly involve multiple data processing steps, such as experimental processing, computational processing, sample selection, or feature selection (i.e. gene selection), prior to deriving any biological conclusions. These steps can dramatically change the interpretation of an experiment. Evaluation of processing steps has received limited attention in the literature. It is not straightforward to evaluate different processing methods and investigators are often unsure of the best method. We present a simple statistical tool, Standardized WithIn class Sum of Squares (SWISS), that allows investigators to compare alternate data processing methods, such as different experimental methods, normalizations, or technologies, on a dataset in terms of how well they cluster a priori biological classes. SWISS uses Euclidean distance to determine which method does a better job of clustering the data elements based on a priori classifications. We apply SWISS to three different gene expression applications. The first application uses four different datasets to compare different experimental methods, normalizations, and gene sets. The second application, using data from the MicroArray Quality Control (MAQC) project, compares different microarray platforms. The third application compares different technologies: a single Agilent two-color microarray versus one lane of RNA-Seq. These applications give an indication of the variety of problems that SWISS can be helpful in solving. The SWISS analysis of one-color versus two-color microarrays provides investigators who use two-color arrays the opportunity to review their results in light of a single-channel analysis, with all of the associated benefits offered by this design. Analysis of the MACQ data shows differential intersite reproducibility by array platform. SWISS also shows that one lane of RNA-Seq clusters data by biological phenotypes as well as a single Agilent two-color microarray.
2010-03-26 | GSE20234 | GEO
Project description:WGS of pannonian and swiss Arabidopsis arenosa populations
Project description:In order to identify the molecular mechanisms controlling preadipocyte commitment we derived new sublines of Swiss 3T3 fibroblasts with varying potential for adipogenesis. Swiss 2, Swiss 8, Swiss 9, and Swiss 19 differentiate into fat cells while Swiss 3, Swiss 5, and Swiss 22 have a lower propensity to differentiate into fat cells. The overall goal was to identify genes whose expression in the fibroblast state correlates with the ultimate potential of the cells to undergo adipogenesis
Project description:Intervention type:DRUG. Intervention1:Huaier, Dose form:GRANULES, Route of administration:ORAL, intended dose regimen:20 to 60/day by either bulk or split for 3 months to extended term if necessary. Control intervention1:None.
Primary outcome(s): For mRNA libraries, focus on mRNA studies. Data analysis includes sequencing data processing and basic sequencing data quality control, prediction of new transcripts, differential expression analysis of genes. Gene Ontology (GO) and the KEGG pathway database are used for annotation and enrichment analysis of up-regulated genes and down-regulated genes.
For small RNA libraries, data analysis includes sequencing data process and sequencing data process QC, small RNA distribution across the genome, rRNA, tRNA, alignment with snRNA and snoRNA, construction of known miRNA expression pattern, prediction New miRNA and Study of their secondary structure Based on the expression pattern of miRNA, we perform not only GO / KEGG annotation and enrichment, but also different expression analysis.. Timepoint:RNA sequencing of 240 blood samples of 80 cases and its analysis, scheduled from June 30, 2022..
Project description:We have employed chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) to analyze changes in chromatin architecture as well as the occupancy of two RNA Polymerase II (RNAPII) isoforms, initiation-competent (RNAPIIS5ph) as well as elongation-competent (RNAPIIS2ph) upon stress induction. We used resting mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway. Serum starved (72 h 0.2% FCS) mouse 3T3 cells were treated with anisomycin (188.5 nM) for 1 h (in duplicates). Untreated, serum-starved cells were used as a control. Isolated chromatin was subjected to immunoprecipitation with the following antibodies: a-H3S28ph, a-H3K9ac, a-H3K4me3, a-RNAPIIS5ph and a-RNAPIIS2ph. Resulting DNA was sequenced using Illumina platforms.