|
| Status |
Public on Apr 20, 2016 |
| Title |
4SVR4 |
| Sample type |
RNA |
| |
|
| Source name |
Cultured iWAT
|
| Organism |
Mus musculus |
| Characteristics |
strain: SV129
cell culture duration (in days): 7
treatment: Rosiglitazone
|
| Treatment protocol |
Treated with 1 uM rosiglitazone maleate (EnzoLifeSciences) and 1 uM norepinephrine (NE; Sigma-Aldrich) from the first day of culture.
|
| Growth protocol |
After isolation from inguinal subcutaneous tissue white adipocyte precursors were cultured in growth medium containing DMEM with 10% (vol/vol) newborn calf serum (Invitrogen), 2.4 nM insulin, 25 ug/ml sodium ascorbate, 10 mM HEPES 4mM glutamine, 50 U/ml penicillin, and 50ug/ml streptomycin. The cells were differentiated and cultured at 37°C in an atmosphere of 5% CO2 in air with 80% humidity
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated by TRIzol (Invitrogen) according to the manufacturer’s protocol. RNA was dissolved in 20ul of RNAse-free water and quantified using a Nanodrop 2000 (NanoDrop technologies).
|
| Label |
biotin
|
| Label protocol |
Ambion’s WT expression kit to produce cDNA. The cDNA was fragmented and labeled with GeneChip WT Terminal labeling kit (Affymetrix). The hybridization of cDNA to exon array was 16h at 45 degrees. The arrays were washed in Affymetrix FS450 wash stations
|
| |
|
| Hybridization protocol |
Arrays were washed, stained and scanned following Affymetrix standard procedures.
|
| Scan protocol |
Affymetrix Gene Chip Scanner
|
| Data processing |
The data was analyzed with affymetrix power tools using the RMA algorithm. The expression measure is given in log base 2 scale
|
| |
|
| Submission date |
May 22, 2014 |
| Last update date |
May 03, 2019 |
| Contact name |
James A Timmons |
| E-mail(s) |
jamie.timmons@gmail.com
|
| Organization name |
Augur Precision Medicine LTD
|
| Street address |
Cambridge
|
| City |
Cambridge |
| ZIP/Postal code |
CB1 |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6096 |
| Series (1) |
| GSE57903 |
Transcriptomics to discover splice variants involved in the browning of adipose tissue |
|
