The DNA replication checkpoint transcriptionally upregulates genes that allow cells to adapt to and survive replication stress. Our results show that, in the fission yeast Schizosaccharomyces pombe, the replication checkpoint regulates the entire G(1)/S transcriptional program by directly regulating MBF, the G(1)/S transcription factor. Instead of initiating a checkpoint-specific transcriptional program, the replication checkpoint targets MBF to maintain the normal G(1)/S transcriptional program ...[more]
Project description:In this study we explored the antiviral gene expression induced in the CNS of MHV-infected mice, by performing whole-genome expression profiling. Three different mouse strains (BALB/c, 129SvEv and 129SvEv IFNAR-/- mice), differing in their susceptibility to infection with MHV, were used.
Project description:BxPc-3, Miapaca-2 and ASPC-1 cell lines were treated with 100 µM NS-398 in DMSO or with DMSO alone (control) for 48 h. Total-RNA from each sample was isolated with the RNeasy kit of Qiagen (Hilden, Germany) according to the manufacturer's protocol. The integrity of the isolated RNA was checked on an Agilent Bioanalyser 2100 using the RNA 6000 Nano Kit (Agilent Technologies, Palo Alto, USA) according to the manufacturer's protocol as recommended by the manufacturer.<br>Microarrays were produced and processed as described before. Human cDNAs representing some 7,000 genes that are highly associated with the occurrence of pancreatic cancer including apoptotic and oncogenic genes, growth factors, angiogenic, cell cycle, metastasis-associated, and housekeeping genes were PCR-amplified using amino-modified M13 universal primers. PCR-products were purified with Multiscreen PCR (Millipore, Schwalbach, Germany), resuspended in spotting solution (TeleChem International, Sunnyvale, USA) and arrayed onto slides with epoxysilane surface (Quantifoil Micro Tools, Jena, Germany). DNA spotting was done with a Micro-Arrayer of Engineering Services Inc. (Virtek's arrayer system, BioRad, Munich, Germany) using SMP3 pins (TeleChem). <br><br>Transcript profiling<br>Fluorescently labelled cDNA samples were prepared from 15 µg total-RNA and Cy3- or Cy5-labelled dCTP was directly incorporated during first-strand synthesis . Hybridisation to the microarrays was done in hybridisation chambers (TeleChem) at 62°C overnight. After washing in 0.1xSSC, fluorescence signals were detected with a confocal ScanArray 5000 scanner (Packard Bioscience, USA) and analysed with GenePix Pro 6 (Axon Instruments, Union City, USA). <br>
Project description:Assess the full impact of estrogen receptor beta on transcription by a full transcriptome analysis of ERb-mediated gene regulation in the SW480 colon cancer cell line. The colon cancer cell line SW480 does not express endogenous ER but is made ERb-expressing by lentiviral transduction of an ERb expression cassette. Introduction of ERb makes it possible to study the role and function of ERb in colon cancer as well as the impact ERb has on its own (in the absence of ERa).
Project description:The aim was to study the effects of Nur77 on the white adipose tissue transcriptome after fasting. For this purpose we performed gene expression profiling of white adipose tissue from Nur77-/- mice and wildtype (Nur77+/+) littermates submitted to prolonged fasting using microarray analysis on >27k elements cDNA microarrays.