|
| Status |
Public on Jun 07, 2016 |
| Title |
GFP 2 |
| Sample type |
RNA |
| |
|
| Source name |
GFP control
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Mouse primary myoblasts
|
| Treatment protocol |
Upon myotube formation (48h after induction of differentiation) we expressed GFP alone or GFP and one of the four PGC-1α isoforms via adenoviral vectors at an MOI of 100. GFP and PGC-1α isoforms are expressed from independent promoters. Adenoviruses expressing the different mouse PGC1α isoforms or GFP have been previously described (3). Twelve hours after transduction, media was replaced with fresh Differentiation Media. Approximately 60 hours post-transduction, cells were harvested in Isol-RNA lysis reagent.
|
| Growth protocol |
During the growth phase, mouse primary myoblasts were plated in collagen-coated plates and maintained in Ham´s F-10 Nutrient Mixture (Gibco), 20% FBS (Sigma), 1% Penicillin/Streptomycin (Gibco) and 2.5 ng/ml basic Fibroblast Growth Factor (Gibco). After achieving 90% confluence, cells were changed to Differentiation Media consisting of DMEM (Gibco), 5% Horse Serum (Gibco), and 1% Penicillin/Streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Approximately 60 hours post-transduction, cells were harvested in Isol-RNA lysis reagent (5PRIME) and purified with Nucleospin RNA II columns (Macherey-Nagel) according to manufacturer’s instructions to achieve optimal purity levels
|
| Label |
biotin
|
| Label protocol |
Total RNA was converted to sense strand cDNA using Ambion WT Expression kit. For fragmentation and labeling WT GeneChip WT target Labeling kit was used.
|
| |
|
| Hybridization protocol |
According to Affymetrix protocol
|
| Scan protocol |
According to Affymetrix protocol
|
| Data processing |
Data were processed through Qlucore and Partek.
|
| |
|
| Submission date |
Nov 27, 2015 |
| Last update date |
Jun 07, 2016 |
| Contact name |
Karin Dahlman-Wright |
| Organization name |
Karolinska Institutet
|
| Department |
Department of Biosciences and Nutrition
|
| Lab |
Medicinaren 25/Neo
|
| Street address |
Blickagången 16
|
| City |
Huddinge |
| ZIP/Postal code |
14157 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL6096 |
| Series (1) |
| GSE75448 |
Peroxisome proliferator-activated receptor gamma coactivator-1alpha isoforms selectively regulate multiple splicing events on target genes. |
|
