|
| Status |
Public on Apr 15, 2011 |
| Title |
control uninephrectomy |
| Sample type |
RNA |
| |
|
| Source name |
pooled RNA from 4 FVB-uni mice
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: FVB/NJ
genotype: control
tissue: kidney
gender: female
age: 4.5 months
disease: nondiabetic
surgery: uninephrectomy
time after surgery: 10 weeks
|
| Treatment protocol |
Uninephrectomy was applied to OVE26 mice and their littermates at the age of 2 months. All mice were sacrificed under anesthesia at 4.5 months of age, which was 10 weeks after surgery. Kidneys were harvested under anesthesia and perfused with phosphate-buffered saline. Half of the kidney from each animal was snap frozen in liquid nitrogen and stored at -80C for RNA extraction.
|
| Growth protocol |
OVE diabetic mice on the FVB background were bred in our animal facility and had free access to standard rodent chow and water throughout the study.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from frozen kidney using TriZol reagent. RNA was pooled from 4 mice in each group (OVE-uni, OVE-sham, FVB-uni, FVB-sham). A 100ng aliquot of RNA from the pooled sample was used for probe preparation.
|
| Label |
biotin
|
| Label protocol |
3' IVT Express Kit was used for biotin labeling.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA was hybridized for 16 hr at 45C on a GeneChip Mouse Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
Arrays were scanned using a GCS 3000 7G scanner and GCOS software (Affymetrix, Santa Clara, CA).
|
| Description |
FVB-uni: uninephrectomy treatment in nondiabetic mice.
|
| Data processing |
To identify genes altered by diabetes, two comparisons of signal intensity were made, OVE-uni versus FVB-uni (designated as UNI ratio) and OVE-sham versus FVB-sham (designated as SHAM ratio). The ratio of normalized signal intensity was determined for each probe set on the chip. Genes with a two-fold or greater change in signal were uploaded to Ingenuity software (Ingenuity Pathway Analysis, Redwood City, CA) for canonical pathway analysis.
|
| |
|
| Submission date |
Apr 21, 2010 |
| Last update date |
Apr 15, 2011 |
| Contact name |
Shirong Zheng |
| E-mail(s) |
[email protected]
|
| Phone |
502-852-2632
|
| Organization name |
University of Louisville
|
| Department |
Pediatrics
|
| Lab |
Dr. Paul N Epstein
|
| Street address |
570 S Preston St.
|
| City |
Louisville |
| State/province |
KY |
| ZIP/Postal code |
40202 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE21448 |
Renal gene expression in uninephrectomized diabetic OVE26 mice |
|
