|
| Status |
Public on Jan 04, 2016 |
| Title |
Insulin treatment, biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
C2C12 cells, insulin
|
| Organism |
Mus musculus |
| Characteristics |
cell line: C2C12
cell type: differentiated myotubes
treatment: chronic insulin
|
| Treatment protocol |
For chronic insulin treatment, cells were either left untreated or incubated with 100 nM insulin in DMEM for 48h in fusion medium to induce an insulin-resistance state. Medium was changed every 24h.
|
| Growth protocol |
Briefly, C2C12 cells were cultured in Dulbecco's Modified Eagle Media (DMEM) supplemented with 10% fetal bovine serum, and penicillin/streptomycin. To induce differentiation, media was replenished by DMEM containing 2% (v/v) of horse serum with penicillin/streptomycin. Myotubes between days 4 and 7 following the induction of differentiation were used for experiments.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with a combination of Trizol and the RNeasy Mini Kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
25 ng total RNA was amplified using the TransPlex® Complete Whole Transcriptome Amplification Kit (Sigma; reference WTA2), and subsequently labeled using the GeneChip Mapping 10K Xba Assay Kit (Affymetrix; catalog # 900441) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
8 µg of cDNA was hybridized per microarray for 16 hours of hybridization at 45ºC. Washing and staining of microarrays were performed using a Fluidics Station 450 (Affymetrix, Santa Clara, CA).
|
| Scan protocol |
GeneChips were scanned in a GeneChip Scanner 3000 (Affymetrix, Santa Clara, CA).
|
| Description |
SMMarch10..347.2010.I3
|
| Data processing |
We applied quantile normalization. For each gene, we selected the 50% of the probes with the highest interquartile-range across samples, and applied RMA summarization to the selected probes. We used the Bioconductor software.
|
| |
|
| Submission date |
Jul 12, 2012 |
| Last update date |
Jan 04, 2016 |
| Contact name |
Josep M Mercader |
| E-mail(s) |
josepmaria.mercader@gmail.com
|
| Phone |
+34661327724
|
| Organization name |
Barcelona Supercomputing Center
|
| Department |
Life Science Department
|
| Lab |
Computational Genomics
|
| Street address |
C/Jordi Girona 29 08034
|
| City |
Barcelona |
| State/province |
Catalunya |
| ZIP/Postal code |
08034 |
| Country |
Spain |
| |
|
| Platform ID |
GPL10740 |
| Series (2) |
| GSE39317 |
Identification of novel type 2 diabetes candidate genes involved in the crosstalk between the mitochondrial and the insulin signaling systems: chronic insulin treatment |
| GSE39320 |
Identification of novel type 2 diabetes candidate genes involved in the crosstalk between the mitochondrial and the insulin signaling systems |
|
