|
| Status |
Public on Mar 29, 2012 |
| Title |
BMM_rep2_24 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse bone marrow-derived macrophages stimulated with 10ng/ml S. minnesota lipopolysaccharide for 24h.
|
| Organism |
Mus musculus |
| Characteristics |
cell type: bone marrow-derived macrophages
lps treatment (time): 24 hours
|
| Treatment protocol |
BMM 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. 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 |
biotin
|
| Label protocol |
Biotin-labelled cRNA. Refer to the Illumina Gene Expression System Manual for details.
|
| |
|
| Hybridization protocol |
Refer to the Illumina Gene Expression System Manual for details.
|
| Scan protocol |
Refer to the Illumina Gene Expression System Manual for details.
|
| Description |
no additional information
|
| Data processing |
The scanned microarrays were processed using Illumina Beadstudio software (no background subtraction or normalisation). Data was normalised using Genespring. Values below 0.01 were set to 0.01. Each measurement was divided by the 50.0th percentile of all measurements in that sample (per-chip normalisation). Specific samples were normalized to one another (per-gene normalisation): all replica one samples were were normalized against the mean of the replica one unstimulated control sample; and all replica two samples were were normalized against the mean of the replica two unstimulated control samples (which were hybridised in technical duplicate ie. the same RNA hybridised twice).
|
| |
|
| Submission date |
Jan 06, 2010 |
| 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 |
GPL4055 |
| Series (2) |
| GSE19492 |
Conservation and divergence in Toll-like receptor 4-regulated gene expression in primary human versus mouse macrophages |
| GSE19766 |
Transcriptional responses of mouse bone marrow-derived macrophages (BMM) to lipopolysaccharide (LPS) - Illumina arrays |
|
