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Sample GSM357355 Query DataSets for GSM357355
Status Public on Apr 01, 2009
Title MNNG biological replicate 4
Sample type RNA
 
Channel 1
Source name MNNG 1μM 24h
Organism Homo sapiens
Characteristics Caco-2 (Human colonic adenocarcinoma cells)
Treatment protocol Caco-2 cells were treated for 24 hours with one of the following six NOC: MNNG (1μM), MNU (1mM), NDEA (50mM), NDMA (100mM), NPIP (40mM) and NPYR (100mM)
Growth protocol Caco-2 cell cultures were transferred weekly by trypsinization and incubated at 37 ºC in a humidified incubator containing 5% CO2
Extracted molecule total RNA
Extraction protocol RNA was isolated from TRIzol® suspended cells according to the manufacturer’s protocol with minor modifications, followed by a clean up, using an RNeasy Mini Kit (Qiagen, Venlo, The Netherlands) with DNase treatment.
Label Cy3
Label protocol The Two-Color Microarray-Based Gene Expression Analysis kit from Agilent Technologies (Amstelveen, The Netherlands) was used to generate Cyanine (Cy) labelled cRNA according to the manufacturer’s protocol.
 
Channel 2
Source name Control for MNNG 24h
Organism Homo sapiens
Characteristics Caco-2 (Human colonic adenocarcinoma cells)
Treatment protocol Caco-2 cells were treated for 24 hours with one of the following six NOC: MNNG (1μM), MNU (1mM), NDEA (50mM), NDMA (100mM), NPIP (40mM) and NPYR (100mM)
Growth protocol Caco-2 cell cultures were transferred weekly by trypsinization and incubated at 37 ºC in a humidified incubator containing 5% CO2
Extracted molecule total RNA
Extraction protocol RNA was isolated from TRIzol® suspended cells according to the manufacturer’s protocol with minor modifications, followed by a clean up, using an RNeasy Mini Kit (Qiagen, Venlo, The Netherlands) with DNase treatment.
Label Cy5
Label protocol The Two-Color Microarray-Based Gene Expression Analysis kit from Agilent Technologies (Amstelveen, The Netherlands) was used to generate Cyanine (Cy) labelled cRNA according to the manufacturer’s protocol.
 
 
Hybridization protocol Hybridization according to Agilent instructions.
Scan protocol Slides were scanned on a GenePix® 4000B Microarray Scanner (Molecular Devices, Sunnyvale, USA). Cy3 and Cy5 were excited at wavelengths of 532 and 635 nm, respectively. Laser power was set to 100%. The photo multiplier tube gain was set to a saturation tolerance of 0.02% to minimize background and saturated spots. Photomultiplier gain was set to 587 (Cy3) and 705 (Cy5) for replicates 1 and 2, and 628 (Cy3) and 560 (Cy5) for replicates 3 and 4, based on automatic establishment of ideal scan settings. The images obtained (resolution 5 micron, 16 bit tiff image) were processed with Imagene 8.0.1 software (Biodiscovery, El Segundo, USA) to measure mean signal intensities for spots and local backgrounds followed by a quality control in Microsoft Excel.
Description Biological replicate 4 of 4
Data processing Data preparations were performed in GeneSight software (BioDiscovery) and included in chronological order: background subtraction, omission of bad spots (controls and irregularly shaped spots), LOWESS normalization, log (base=2) transformations, calculation of difference of test compound versus respective vehicle control.
 
Submission date Jan 05, 2009
Last update date Jan 05, 2009
Contact name Dennie Hebels
E-mail(s) d.hebels@maastrichtuniversity.nl
Phone 0031-43-3881088
Fax 0031-43-3884146
URL http://www.grat.nl
Organization name Maastricht University
Department Health Risk Analysis and Toxicology
Street address Universiteitssingel 50
City Maastricht
ZIP/Postal code 6200MD
Country Netherlands
 
Platform ID GPL6480
Series (1)
GSE14284 Molecular signatures of N-nitroso compounds in Caco-2 cells: implications for colon carcinogenesis

Data table header descriptions
ID_REF
VALUE Background corrected, LOWESS normalized log2 ratio (test compound/control)

Data table
ID_REF VALUE
A_23_P100001 0.3520
A_23_P100011 0.3793
A_23_P100022
A_23_P100056 0.5704
A_23_P100074 -0.0034
A_23_P100092 -0.0216
A_23_P100103 0.3653
A_23_P100111 0.4900
A_23_P100127 0.0412
A_23_P100133 0.3595
A_23_P100141 0.0613
A_23_P100156 -0.0687
A_23_P100177 0.8241
A_23_P100189 0.1642
A_23_P100196 0.3675
A_23_P100203 -0.0868
A_23_P100220 -0.0905
A_23_P100240 0.0361
A_23_P10025
A_23_P100263 0.1450

Total number of rows: 41000

Table truncated, full table size 736 Kbytes.




Supplementary file Size Download File type/resource
GSM357355_Cy3.txt.gz 4.2 Mb (ftp)(http) TXT
GSM357355_Cy5.txt.gz 4.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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