|
| Status |
Public on Sep 13, 2016 |
| Title |
Control _vs_growth hormone-treated 3T3-F442A adipocytes |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
3T3 cell line_control
|
| Organism |
Mus musculus |
| Characteristics |
cell line: 3T3-F442A
cell type: adipocytes
|
| Treatment protocol |
The differentiated 3T3-F442A cells were treated with GH for 24 H (at final concentration of 125 ng/mL).
|
| Growth protocol |
The mouse embryo fibroblasts 3T3-F442A were cultured and induced to differentiate, at 12 days approximately 80% 80% 3T3-F442A cells were differentiated into adipocytes. And then cultures with DMEM containing 10% FBS every other day.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction. This procedure has been previously described, 2 and the procedure has been improved by using CapitalBio cRNA Amplification and Labeling Kit (CapitalBio) for producing higher yields of labeled cDNA.
|
| |
|
| Channel 2 |
| Source name |
3T3 cell line_growth hormone-treated
|
| Organism |
Mus musculus |
| Characteristics |
cell line: 3T3-F442A
cell type: adipocytes
|
| Treatment protocol |
The differentiated 3T3-F442A cells were treated with GH for 24 H (at final concentration of 125 ng/mL).
|
| Growth protocol |
The mouse embryo fibroblasts 3T3-F442A were cultured and induced to differentiate, at 12 days approximately 80% 80% 3T3-F442A cells were differentiated into adipocytes. And then cultures with DMEM containing 10% FBS every other day.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction. This procedure has been previously described, 2 and the procedure has been improved by using CapitalBio cRNA Amplification and Labeling Kit (CapitalBio) for producing higher yields of labeled cDNA.
|
| |
|
| |
| Hybridization protocol |
DNA in hybridization solution was denatured at 95℃ for 3 min prior to loading onto a microarray. Arrays were hybridized was preformed in a CapitalBio BioMixerTM II Hybridization Station overnight at a rotation speed of 8 rpm and a temperature of 42℃ and washed with two consecutive solutions (0.2% SDS, 2× SSC at 42℃ for 5 min, and 0.2× SSC for 5 min at room temperature).
|
| Scan protocol |
Arrays were scanned with a confocal LuxScanTM scanner and the images obtained were then analyzed using LuxScanTM 3.0 software (Both from CapitalBio)
|
| Data processing |
LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal.
|
| |
|
| Submission date |
Sep 12, 2016 |
| Last update date |
Sep 13, 2016 |
| Contact name |
Yuchao Zhang |
| E-mail(s) |
yc.zhang@hotmail.com
|
| Organization name |
Shandong University
|
| Street address |
44 Wenhua Road West
|
| City |
Jinan |
| ZIP/Postal code |
250012 |
| Country |
China |
| |
|
| Platform ID |
GPL22425 |
| Series (1) |
| GSE86830 |
Murine 3T3-F442A Cells: Control vs. Growth hormone |
|
