|
| Status |
Public on Oct 22, 2020 |
| Title |
LPS injected healthy control, biological rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Whole brain RNA, 48hr after LPS injection (i.p.)
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL6
Sex: Male
tissue: Brain
|
| Treatment protocol |
Male C57BL6 mice were injected (i.p.) with 4mg/kg LPS (Escherichia coli serotype 055:B5) or PBS (n=3/group).
|
| Growth protocol |
C57BL/6 mice (7 to 8 weeks old, 20 to 25 g) were purchased from Harlan Laboratories. Mice were maintained in specific pathogen-free conditions in standard caging in the Central Research Facility at the University of Glasgow. Mice were givin 1 week to acclimatise prior to treatment.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Whole brain tissue was snap frozen and stored at -80°C until use. Under RNase-free conditions, brains were homogenized using the TissueLyser LT (Qiagen, Hilden, Germany). RNA was extracted from homogenized tissue using Trizol® (Life Technologies, Invitrogen, Carlsbad, CA, USA) as described by the manufacturers. Isolated RNA was further purified and genomic DNA removed using an RNeasy Mini Kit (Qiagen, Hilden, Germany). Red blood cells were lysed from blood samples using red blood cell lysis buffer (Miltenyi, Cologne, Germany). Under RNase-free conditions, RNA was isolated and genomic DNA was removed from PBLs using an RNeasy Micro Kit (Qiagen, Hilden, Germany).
|
| Label |
Biotin
|
| Label protocol |
cDNA fragmentation and labelling was performed using a GeneChip WT Terminal Labelling kit (Affymetrix, Santa Clara, CA, USA).
|
| |
|
| Hybridization protocol |
Fragmented cDNA samples were then hybridized to GeneChip Mouse Gene 1.0 ST Arrays using GeneTitan hybridization
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G
|
| Description |
Gene expression in the brain, 48 hours after the induction of systemic inflammation by LPS injection
|
| Data processing |
Data generated using GeneSpring GX software were normalized using RMA16. The normalized data were subsequently analyzed using t-test to determine the significance of each gene in LPS-treated mice compared with vehicle-treated controls.
|
| |
|
| Submission date |
Oct 21, 2020 |
| Last update date |
Oct 22, 2020 |
| Contact name |
Jonathan Cavanagh |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Glasgow
|
| Department |
Immunology
|
| Street address |
120 University Place
|
| City |
Glasgow |
| ZIP/Postal code |
G12 0TA |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE159770 |
Peripheral inflammation is associated with remote global gene expression changes in the brain |
|
