|
| Status |
Public on Mar 29, 2012 |
| Title |
TEPM-6h LPS-rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse thioglycollate-elicited peritoneal macrophages stimulated with 10ng/ml S. minnesota lipopolysaccharide for 6h
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6
gender: male
tissue: peritoneal
cell type: macrophages
treatment: LPS
treatment time: 6 hours
|
| Treatment protocol |
BMM and TEPM were treated for various times with lipopolysaccharide (LPS) from Salmonella minnesota (Sigma-Aldrich), at a final concentration of 10 ng/ml.
|
| Growth protocol |
BMM were derived from femurs of 6-8 week-old, male C57Bl/6 mice. Femurs were flushed with medium and bone marrow cells were plated out in complete medium (RPMI 1640 containing 10% FCS, 20U/ml penicillin, 20 µg/ml streptomycin and 2mM L-glutamine) supplemented with 104U/ml (100ng/ml) recombinant human CSF-1 (Chiron) on bacteriological plastic plates (Bibby Sterilin) for 7 days, with refeeding on day 5 and reseeding on day 6 at 1x105 cells/cm2. TEPM were elicited by injecting 6-8 week-old male, mice with 1ml sterile thioglycollate broth (10% in pyrogen-free water, Becton Dickinson) intraperitoneally. 5 days post-injection, peritoneal cells were collected by PBS lavage, plated on tissue culture plastic as for BMM in media without CSF-1 and stimulated the following day. >90% TEPM were F4/80+ macrophages, as assessed by surface staining and flow cytometry (not shown). All cells were cultured at 37˚C with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using the Qiagen RNeasy Mini kit with on-column DNaseI treatment, according to the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Labelling was performed using the Agilent Quick-Amp Labelling Kit (one colour) according to manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Samples were hybridised according to Agilent's One-colour Microarray-Based Gene Expression Analysis (Quick Amp Labelling) protocol, V5.7 March 2008.
|
| Scan protocol |
Samples were scanned according to Agilent's One-colour Microarray-Based Gene Expression Analysis (Quick Amp Labelling) protocol, V5.7 March 2008, using an Agilent Microarray Scanner G2565BA.
|
| Description |
TEPM-6h LPS-rep3
|
| Data processing |
Features (including processed signal and flags) were extracted using the Feature Extraction 9.5.3 (Agilent Technologies) software, with default settings and no background subtraction. Processed signal intensities were quantile normalized in R using the Affymetrix package in Bioconductor.
|
| |
|
| Submission date |
Dec 15, 2009 |
| Last update date |
Mar 29, 2012 |
| Contact name |
Kate Schroder |
| E-mail(s) |
K.Schroder@imb.uq.edu.au
|
| Phone |
+617 33462087
|
| Fax |
+617 33462101
|
| Organization name |
Institute for Molecular Bioscience
|
| Street address |
University of Queensland, St Lucia
|
| City |
Brisbane |
| State/province |
QLD |
| ZIP/Postal code |
4072 |
| Country |
Australia |
| |
|
| Platform ID |
GPL9802 |
| Series (2) |
| GSE19490 |
Transcriptional responses of mouse BMM and TEPM to lipopolysaccharide (LPS) |
| GSE19492 |
Conservation and divergence in Toll-like receptor 4-regulated gene expression in primary human versus mouse macrophages |
|
