|
| Status |
Public on Mar 07, 2017 |
| Title |
Dystrophic mouse quadriceps, spironolactone plus lisinopril-treated, biological replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
quadriceps, utrn+/-;mdx, spironolactone plus lisinopril-treated
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: C57BL/10
genotype/variation: utrn+/-;mdx
age: 6 weeks
Sex: male
tissue: quadriceps muscle
treatment: spironolactone plus lisinopril
|
| Treatment protocol |
20 mg/kg x day lisinopril plus 100 mg/kg x day spironolactone delivered via medicated pellets from 4-6 weeks-of-age, 20 mg/kg x day lisinopril plus 200 mg/kg x day eplerenone delivered via medicated pellets from 4-6 weeks-of-age, 1.5 mg/kg x day prednisolone delivered via water bottle from 4-6 weeks-of-age or untreated normal chow.
|
| Growth protocol |
Mouse quadriceps muscles dissected from 6-week-old dystrophic mice.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using TRIzol reagent (Life Technologies), according to manufacturer’s instructions. RNA concentration was determined spectrophotometrically using a Nanodrop ND-100 (Nanodrop Technologies). Samples were DNase treated using RQ1 DNase (Promega) for 30 minutes at 37⁰C, reaction was stopped with a 25:24:1 phenol: chloroform: isoamyl alcohol mixture (Malinckrodt, Fischer, Sigma) and centrifuged 5 minutes (14,000 x g; room temperature). A chloroform phase separation was performed; the aqueous layer was removed and ethanol (Fisher) precipitated. The pellet was washed (75% ethanol), centrifuged (14,000 x g, 10 minutes, 4⁰C), air dried (5 minutes) and re-suspended in autoclaved Millipore filtered water. DNase treated RNA concentration was determined spectrophotometrically and samples were stored at -80⁰C. Samples for microarray analysis were further purified using the RNeasy mini kit (Qiagen) RNA cleanup protocol and final RNA concentrations were determined. RNA integrity was interrogated using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). A 100 ng aliquot of total mRNA was linearly amplified.
|
| Label |
biotin
|
| Label protocol |
5.5 µg of cDNA was labeled and fragmented using the GeneChip WT PLUS reagent kit (Affymetrix) following the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Labeled cDNA targets were hybridized to Affymetrix GeneChip® Mouse Transcriptome array 1.0 for 16 h at 45 ⁰C rotating at 60 rpm.
|
| Scan protocol |
The arrays were washed and stained using the Fluidics Station 450 and scanned using the GeneChip Scanner 3000.
|
| Description |
6174
|
| Data processing |
For gene expression analysis, arrays were normalized using Gene Level SST RMA algorithm in Expression Console and comparisons made in Transcriptome Analysis Console (Affymetrix).
|
| |
|
| Submission date |
Jul 26, 2016 |
| Last update date |
May 05, 2017 |
| Contact name |
Jill Rafael-Fortney |
| E-mail(s) |
[email protected]
|
| Organization name |
The Ohio State University
|
| Department |
Physiology & Cell Biology
|
| Lab |
390 Biomedical Research Tower
|
| Street address |
460 W. 12th Ave
|
| City |
Columbus |
| State/province |
OH |
| ZIP/Postal code |
43210 |
| Country |
USA |
| |
|
| Platform ID |
GPL20258 |
| Series (1) |
| GSE84876 |
Expression data from quadriceps of utrn+/-;mdx mice treated with spironolactone plus lisinopril, eplerenone plus lisinopril or prednisolone compared to untreated |
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