|
| Status |
Public on Aug 01, 2014 |
| Title |
PEC-enriched cells isolate replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Capsulated glomeruli
|
| Organism |
Rattus norvegicus |
| Characteristics |
age: Adult
gender: male
strain: Sprague-Dawley
|
| Treatment protocol |
After discarding medulla and papilla, minced cortices were passed through differential stainless steel sieves. In order to obtain capsulated glomeruli enriched in PECs, minced cortices were first pressed through a 180um-sized mesh sieve and then washed through sequential sieves with sizes of 106um, 90um. In order to obtain decapsulated glomeruli deprived of PECs, the 90μm sieve was removed and decapsulated glomeruli were collected on the 75μm sieve. Glomeruli were centrifuged at 80G at 4oC and quantification of glomeruli was performed in aliquots of the glomerular suspension by phase microscopy.
|
| Growth protocol |
No culturing of cells was performed
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from capsulated and decapsulated glomeruli using TRIzol protocol per manufacturer's instructions. RNA quality was confirmed by a Bioanalyzer 2100.
|
| Label |
Biotin
|
| Label protocol |
Biotin labeled cRNA.
|
| |
|
| Hybridization protocol |
Labeled RNA was processed for hybridization to Affymetrix Rat GeneChip 1.0ST microarrays at University of Washington’s Center for Array Technology according to the manufacturer’s instructions.
|
| Scan protocol |
Microarrays were scanned using Affymetrix GeneChip Scanner, and the GeneChip Operating Software (GCOS) was used for image analysis.
|
| Data processing |
Background adjustment and quantile normalization using the Robust Multichip Average algorithm (http://www.bioconductor.org) using Log2 transformed intensities.
|
| |
|
| Submission date |
Jan 15, 2014 |
| Last update date |
Oct 19, 2022 |
| Contact name |
Sina Gharib |
| E-mail(s) |
sagharib@uw.edu
|
| Organization name |
University of Washington
|
| Department |
Medicine
|
| Street address |
850 Republican St.
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98109 |
| Country |
USA |
| |
|
| Platform ID |
GPL6247 |
| Series (1) |
| GSE54085 |
Gene expression analysis of rat glomerular parietal epithelial cells |
|
