biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsIain Dougalltranscription profiling by arrayHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-GEOD-30660Airway epithelial cells are the first cells of the lungs to be exposed to the toxic agents contained within cigarette smoke. Accordingly, the response of these cells to this challenge is of considerable interest in the context of diseases in which cigarette smoke is a major aetiological factor. We used Affymetrix microarrays to study the effects of repeated cigarette smoke challenge on three-dimensional human lung airway epithelial cultures. We have used cultures from four individual donors, and compared the effects of repeated cigarette smoke challenge (30mins exposure on four separate days), with no challenge.biostudies-arrayexpressLabeling - Biotinylated cRNA were prepared according to the standard Affymetrix protocol from total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).Growth Protocol - ALI cultures were maintained in proprietary MatTek mediaHybridization - Following fragmentation, cRNA was hybridized for on HGU133 plus2 human Affymetrix GeneChips according to standard protocols.Sample Processing - Cultures were placed in smoking chambers with basal surface in contact with media. Cutures were subsequently exposed to smoke at a constant smoke/air dilution of 1:10 or 1:20. The cells were continually exposed to smoke (from filtered Kentucky reference (3R4F) cigarettes) for 30mins on days 1, 2, 5 and 6.Nucleic Acid Extraction - RNA was isolated using an RNAqueous-4PCR kit(Ambion AM1914).MIAME ScoreRaw DataOrganizationAssays and DataProcessed DataMAGE-TAB FilesArray DesignsStuart BallMartyn FosterNicholas CoxSally StinsonEdward AinscowMatthew BladesRebecca WallsWendy MerrisonIain DougallAssay Data Transformation - ID_REF = <br>VALUE = log2 RMA signalImage Adquisition - GeneChips were scanned using the Affymetrix GeneChip Scanner 3000Feature Extraction - The data were normalised using the Robust Multichip Average (RMA) algorithm in Bioconductor run in R v2.4.0. Statistical analyses were carried out in SAS v 9.2.falseThe Effect of Repeated Whole Cigarette Smoke Challenge on Human Air-Liquid Interface Lung Epithelial CulturesAirway epithelial cells are the first cells of the lungs to be exposed to the toxic agents contained within cigarette smoke. Accordingly, the response of these cells to this challenge is of considerable interest in the context of diseases in which cigarette smoke is a major aetiological factor. We used Affymetrix microarrays to study the effects of repeated cigarette smoke challenge on three-dimensional human lung airway epithelial cultures. We have used cultures from four individual donors, and compared the effects of repeated cigarette smoke challenge (30mins exposure on four separate days), with no challenge.2011-08-30T00:00:00Z2023-08-13T11:21:57.782Z2022-02-08T00:20:13.864ZE-GEOD-30660GSE30660EFO_0002768