|
| Status |
Public on Jul 09, 2020 |
| Title |
CNF1_D03_1.0mg/kg CNF1 |
| Sample type |
RNA |
| |
|
| Source name |
3 day post-instillation in 1.0mg/kg CNF1-exposed rat lung
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague-Dawley (Crl:CD[SD])
Sex: male
treatment: 1.0mg/kg CNF1
time: Day3
tissue: lung
|
| Treatment protocol |
0.5, 1.0 and 2.0 mg/mL of CNF (CNF1, CNF2, and CNF3) and MWCNT suspensions were prepared with distilled water with preservatives as each vehicle control. Each rat received a single intratracheal instillation dose (1 mg/kg) of the CNFs, MWCNT, and vehicles. Rats were dissected at 1, 3, 7, 30, and 90 days post-instillation. Lungs were perfused with physiological saline, excised, and used for whole genome microarray analysis.
|
| Growth protocol |
Seven-week-old male Sprague-Dawley (Crl:CD[SD]) rats were purchased from Charles River Laboratories Japan (Yokohama, Japan). Rats were housed in metal cages with in a room with 35%– 70% humidity and a controlled temperature of 20°C–26°C and fed a chow diet ad libitum. All procedures and animal handling were approved by the Institutional Animal Care and Use Committee of the National Institute of Advanced Industrial Science (Tsukuba, Japan) and Technology and the BioSafety Research Center Inc. (Iwata, Japan)..
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Blood-perfused rat right lungs (n = 4 per group per time point) were homogenized using QIAzol lysis reagent with a Tissue Ruptor (Qiagen, Tokyo, Japan). Total RNA from the homogenates was extracted using the RNeasy Midi kit (Qiagen) following the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3-labeled cRNA was prepared from RNA using a Low Input Quick Amp Labeling kit (Agilent) according to the manufacturer’s instructions, followed by RNeasy column purification (Qiagen). Dye incorporation and cRNA yield were checked with the NanoDrop 2000 spectrophotometer.
|
| |
|
| Hybridization protocol |
Each labeled cRNA probe was used separately for hybridization to a 4×44 K Whole Rat Genome Microarray (G4131F; Agilent Technologies) at 65 °C for 17 h.
|
| Scan protocol |
Hybridized microarray slides were washed according to the manufacturer’s instructions and scanned with a DNA Microarray Scanner (Agilent, G2505C) at 5 μm resolution.
|
| Description |
Rat lung, Day03, 1.0mg/kg CNF1, replicate 4
|
| Data processing |
The scanned images were analyzed numerically using Agilent Feature Extraction Software, version 12.0.3.1.
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| |
|
| Submission date |
Jul 08, 2020 |
| Last update date |
Jul 09, 2020 |
| Contact name |
Katsuhide Fujita |
| E-mail(s) |
ka-fujita@aist.go.jp
|
| Organization name |
National Institute of Advanced Industrial Science and Technology
|
| Department |
Research Institute of Science for Safety and Sustainability
|
| Street address |
Onogawa 16-1
|
| City |
Tsukuba |
| ZIP/Postal code |
3058569 |
| Country |
Japan |
| |
|
| Platform ID |
GPL7294 |
| Series (1) |
| GSE154013 |
Gene expression profiles in rat lung following intratracheal instillation with cellulose nanofibrils and multi-walled carbon nanotubes |
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