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Series GSE2032 Query DataSets for GSE2032
Status Public on Dec 03, 2004
Title Cardiac cells after mechanical stress, ionising radiation or UV
Organism Rattus norvegicus
Experiment type Expression profiling by array
Summary Rationale: During excessive pressure or overload, cardiac cells are subjected to increased mechanical stress. We investigate how the stress response of cardiac cells to mechanical stress can be compared to genotoxic stresses induced by DNA damaging agents.
Methods: Cultures enriched for cardiomyocytes and cultures of fibroblasts were derived from ventricles of neonatal rat hearts. In three independent experiments, both cell cultures were subjected to mechanical stress (cyclic stretch at 60 cycles/min), UV irradiation (10 J/m2), or X-rays (8.5 Gy). Cells were harvested 24 h. after (the onset of) treatment. Total RNA was isolated using an RNeasy® kit (Qiagen). Single stranded cDNA was synthesized from RNA from myocyte-enriched populations (10-14 microgram) and populations of fibroblasts (7.5-14 microgram), using Superscript II reverse transcriptase and T7-oligo(dT)24 primers at 42ºC for 1 h. Double stranded cDNA was obtained by using DNA ligase, DNA polymerase I and RNAse H at 16ºC for 2 h, followed by T4 DNA polymerase at 16ºC for 5 min. cRNA was synthesized from cDNA using the BioArray HighYield® RNA transcript labeling kit (Enzo) in the presence of biotin-labeled CTP and UTP. cRNA was fragmented in a buffer containing 40 mM Tris-acetate (pH 8.1), 100 mM KAc and 30 mM MgAc, at 94ºC for 35 min. Fragmented cRNA was hybridized to GeneChip RG-U34A arrays (Affymetrix) at 45ºC for 16 h. Arrays were washed and incubated with a streptavidin-phycoerythrin complex. Fluorescent signals were determined with a GeneChip scanner.
Keywords: other
 
 
Contributor(s) Boerma M, Van der Wees CG, Vrieling H, Svensson P, Wondergem J, Van der Laarse A, Mullenders LH, Van Zeeland AA
Citation(s) 15656902
Submission date Dec 02, 2004
Last update date Feb 21, 2017
Contact name Marjan Boerma
E-mail(s) mboerma@uams.edu
Phone 501-686-6882
Organization name University of Arkansas for Medical Sciences
Street address 4301 West Markham, Slot 725
City Little Rock
State/province AR
ZIP/Postal code 72205
Country USA
 
Platforms (1)
GPL85 [RG_U34A] Affymetrix Rat Genome U34 Array
Samples (35)
GSM36457 Fibroblasts ionising radiation 0 Gy
GSM36458 Fibroblasts 24 hours after 8.5 Gy
GSM36459 Cardiac cells 24 hours after sham-irradiation
Relations
BioProject PRJNA91855

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE2032_RAW.tar 67.6 Mb (http)(custom) TAR (of CEL, EXP)

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