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Sample GSM1488937 Query DataSets for GSM1488937
Status Public on Aug 20, 2015
Title NRCWE-030.28.54 Sample 19
Sample type RNA
 
Channel 1
Source name Mouse lung tisue, middle dose, day28
Organism Mus musculus
Characteristics sample id: 97
genetic background: C57BL/6
tissue: lung
dose: middle dose
time: day28
treatment: NRCWE-030
Treatment protocol each mouse received 18, 54, or 162 µg of TiO2NPs or only the vehicle via single intratracheal instillation. Mice were anaesthetised and intratracheally instilled with particle suspensions as described previously (Saber et al 2012, Particle and Fiber Toxicology). Whole lung tissues were collected 24 h and 28 d post exposure, flash frozen in liquid nitrogen and stored at -80°C until the analysis.
Extracted molecule total RNA
Extraction protocol In brief, a small frozen section of lung tissue was homogenized in Trizol (Invitrogen, Carlsbad, CA, USA) using the Retsch Mixer MM 400. RNA was isolated using chloroform, precipitated using isopropyl alcohol and purified using RNeasy Mini Kits (Qiagen, Mississauga, ON, Canada). All RNA showed high integrity with a A260/280 ratio between 2.0 and 2.2 and RNA integrity number above 7.0.
Label Cy5
Label protocol Agilent Linear Amplification Kit (Agilent Technologies Inc., Mississauga, ON, Canada) was used to synthesize cDNA and labelled cRNA from 200 ng of total RNA derived from each individual mouse lungs and commercially available Universal Mouse Reference RNA (UMRR, Stratagene, Mississauga, ON, Canada). Cyanine-labelled cRNA was in vitro transcribed using T7 RNA polymerase and purified using RNeasy Mini Kits (Qiagen, Mississauga, ON, Canada). Experimental samples were labelled with Cyanine-5 and the UMRR was labelled with Cyanine-3.
 
Channel 2
Source name Universal Mouse Reference RNA (UMRR, Stratagene, Mississauga, ON, Canada)
Organism Mus musculus
Characteristics sample type: Universal Mouse Reference RNA (UMRR, Stratagene, Mississauga, ON, Canada)
Treatment protocol each mouse received 18, 54, or 162 µg of TiO2NPs or only the vehicle via single intratracheal instillation. Mice were anaesthetised and intratracheally instilled with particle suspensions as described previously (Saber et al 2012, Particle and Fiber Toxicology). Whole lung tissues were collected 24 h and 28 d post exposure, flash frozen in liquid nitrogen and stored at -80°C until the analysis.
Extracted molecule total RNA
Extraction protocol In brief, a small frozen section of lung tissue was homogenized in Trizol (Invitrogen, Carlsbad, CA, USA) using the Retsch Mixer MM 400. RNA was isolated using chloroform, precipitated using isopropyl alcohol and purified using RNeasy Mini Kits (Qiagen, Mississauga, ON, Canada). All RNA showed high integrity with a A260/280 ratio between 2.0 and 2.2 and RNA integrity number above 7.0.
Label Cy3
Label protocol Agilent Linear Amplification Kit (Agilent Technologies Inc., Mississauga, ON, Canada) was used to synthesize cDNA and labelled cRNA from 200 ng of total RNA derived from each individual mouse lungs and commercially available Universal Mouse Reference RNA (UMRR, Stratagene, Mississauga, ON, Canada). Cyanine-labelled cRNA was in vitro transcribed using T7 RNA polymerase and purified using RNeasy Mini Kits (Qiagen, Mississauga, ON, Canada). Experimental samples were labelled with Cyanine-5 and the UMRR was labelled with Cyanine-3.
 
 
Hybridization protocol Equal amount (300 ng) of labelled cRNA from each experimental sample was hybridized to Agilent Sureprint G3 Mouse GE 4x44K microarrays (agilent Technologies Inc., Mississuaga, ON, Canada) at 65°C for 17 hours in the Agilent SureHyb Hybridization chamber.
Scan protocol Scanned on an Agilent G2565AA scanner.
Data processing Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1). Non background subtracted median signal intensities were LOWESS normalized using the maanova library in R. Probes with technical replicates were averaged using the median.
 
Submission date Aug 26, 2014
Last update date Aug 20, 2015
Contact name Andrew Williams
Organization name Health Canada
Street address 50 Colombine Dr
City Ottawa
State/province ON
ZIP/Postal code K1A0K9
Country Canada
 
Platform ID GPL7202
Series (2)
GSE60799 Transcriptional profiling to identify physical-chemical properties detrimental to nanomaterial-induced pulmonary response (part 3)
GSE60801 Transcriptional profiling to identify physical-chemical properties detrimental to nanomaterial-induced pulmonary response

Data table header descriptions
ID_REF
VALUE Normalized log2 ratio (Cy5/Cy3) representing Sample/reference

Data table
ID_REF VALUE
A_52_P616356 -0.079631289
A_52_P403405 0.14164044
A_52_P819156 0.102317157
A_51_P331831 1.781884233
A_51_P430630 0.196737425
A_52_P502357 0.072102072
A_52_P299964 0.230774552
A_51_P356389 0.238827067
A_52_P684402 -0.967601738
A_51_P414208 0.265415743
A_51_P280918 -0.063666799
A_52_P613688 0.158272828
A_52_P258194 -0.225144994
A_52_P229271 -0.063333099
A_52_P579519 -0.223757983
A_52_P979997 0.030318452
A_52_P453864 0.52497152
A_52_P655842 0.182962232
A_51_P282374 -0.182634342
A_52_P176013 0.262530701

Total number of rows: 41174

Table truncated, full table size 1019 Kbytes.




Supplementary file Size Download File type/resource
GSM1488937_251486837618_201301161152_S01_GE2_1100_Jul11_1_2.txt.gz 4.3 Mb (ftp)(http) TXT
Processed data included within Sample table

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