|
| Status |
Public on Oct 30, 2013 |
| Title |
wt-RLN-78-weeks-1 |
| Sample type |
RNA |
| |
|
| Source name |
Lymph node renal
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Lymph node renal
strain: C57BL/6J
gender: male
age: 78 weeks
|
| Treatment protocol |
The abdominal aorta was exposed, perfused in situ with ice-cold PBS/2mM EDTA and was removed inclusive of paraaortal lymph nodes, adipose and connective tissue. The isolated abdominal aorta was cut in half and embedded in Tissue Tec (Sakura Finetek, Zoeterwoude, NL) and was frozen at -80 centigrade. 200 unstained 10 micrometer cryo-sections of the associated renal lymph node were microdissected with a Palm MicroBeam laser system (P.A.L.M. Microlaser Technologies AG, Bernried, Germany) and pooled in Trizol (Invitrogen, Carlsbad, CA)(according to published procedures: Laser-Capture Microdissection of Hyperlipidemic/ApoE-/- Mouse Aorta Atherosclerosis. Methods Mol Biol. 2011;755:417-28).
|
| Growth protocol |
ApoE-/- and wild-type mice on the C57BL/6J background were from The Jackson Laboratories (Bar Harbor, Maine). Mice were housed in SPF conditions with standard chow diet and were euthanized by carbon dioxide inhalation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using a phenol/chloroform extraction according to the manufacturer's structions. As a last step of the extraction procedure, the RNA was purified with the RNeasy Micro-kit (Qiagen, Hilden, Germany).
|
| Label |
Biotin/SAPE with Anti Streptavidin signal amplification
|
| Label protocol |
T7-High-Yield-RNA-Transcript-Labeling-Kit (Enzo, Farmingdale, NY) with an IVT of 68h
|
| |
|
| Hybridization protocol |
60rpm, 18h, 45 centigrade in Affymetrix Hybridization Buffer
|
| Scan protocol |
Affymetrix Scanner 3000, single scan
|
| Description |
Staining with Fluidic Station 450 Protocol: Midi-euk2v3_450
|
| Data processing |
GCOS 1.4, scaled to 500
|
| |
|
| Submission date |
Aug 06, 2012 |
| Last update date |
Oct 31, 2013 |
| Contact name |
Andreas Johann Richard Habenicht |
| E-mail(s) |
Andreas.habenicht@med.uni-muenchen.de
|
| Organization name |
Ludwig Maximilians Universität München
|
| Department |
Medicine
|
| Lab |
Institute for Cardiovascular Disease Prevention
|
| Street address |
Pettenkoferstraße 9a
|
| City |
Munich |
| State/province |
Bavaria |
| ZIP/Postal code |
80336 |
| Country |
Germany |
| |
|
| Platform ID |
GPL8321 |
| Series (1) |
| GSE40156 |
Transcript atlases reveal that artery tertiary lymphoid organs but not secondary lymphoid organs control key steps of atherosclerosis T cell immunity in aged apoe-/- mice. |
|
