|
| Status |
Public on Mar 28, 2022 |
| Title |
WT MB 1 |
| Sample type |
RNA |
| |
|
| Source name |
Proliferating myoblasts
|
| Organism |
Mus musculus |
| Characteristics |
cell type: myoblasts
genotype: wild type
|
| Growth protocol |
Conditionally immortalized control C57BL/6 and Capn3-deficient myoblast clones contain a temperature-sensitive immortalizing gene, the tsA58 mutation of the SV40 large T antigen, under the control of the H-2Kb inducible promoter. When induced with interferon-gamma (IFN-γ), the tsA58 protein is expressed, allowing the cells to proliferate at 33 °C. Myoblasts were plated and maintained on gelatin-coated dishes (0.01% gelatin) in H2KPM at 33 °C and 10% CO2 in a humidified incubator. Cultures were fed every 2–3 days with H2KPM, and fresh IFN-γ (20 U/ml) was added to the culture medium at each feeding. Cells were grown to near confluence, harvested with trypsin/EDTA (0.5/0.2 g/l), and centrifuged at 800g for 3 min. Under these conditions, the cells doubled every 24–30 h at 33 °C and fused with > 50% efficiency into multinucleated sarcomeric tropomyosin-positive muscle fibers when switched to 37 °C in the absence of IFN-γ.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with TRIzol Reagent followed by clean-up with QIAGEN RNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyzer.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared from 250 ng of total RNA using an Illumina Totalprep RNA amplification kit according to manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
replicate 1
|
| Data processing |
data were extracted using Genome Studio (software provided by Illumina), with no background subtraction, then normalized using the “quantiles” function.
Processed data are available only as fold-change values and p-values.
|
| |
|
| Submission date |
Mar 28, 2022 |
| Last update date |
Mar 28, 2022 |
| Contact name |
Jennifer M Peterson |
| E-mail(s) |
jennifer.peterson@utoledo.edu
|
| Organization name |
The University of Toledo
|
| Department |
SERS
|
| Street address |
2801 W Bancroft St
|
| City |
Toledo |
| State/province |
OH |
| ZIP/Postal code |
43606 |
| Country |
USA |
| |
|
| Platform ID |
GPL6887 |
| Series (1) |
| GSE199620 |
Mitochondrial dysfunction and consequences in calpain-3-deficient muscle myoblasts |
|
