|
| Status |
Public on Apr 09, 2013 |
| Title |
KO_PMΦ_4hLPS_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Nfkbiz-/- peritonial macrophages incubated with 1µg/ml LPS for 4h
|
| Organism |
Mus musculus |
| Characteristics |
gender: female
cell type: peritoneal macrophages
strain: Nfkbiz-/-
|
| Treatment protocol |
Cells were treated with 1 µg/mL LPS for 4 h or left untreated
|
| Growth protocol |
Female C57Bl/6 mice were euthanized by CO2 asphyxiation. For isolation of peritoneal macrophages (PMφ) the abdominal skin was removed, a catheter (24 GA) was inserted into the peritoneal cavity and 10 mL of ice-cold PBS were injected. After massage of the peritoneum, peritoneal fluids were aspirated, centrifuged at 500 g for 5 min, and yielded peritoneal cells (PMφ were resuspended in 0.5-1.5 mL Mφ medium containing DMEM, 10% FCS and MycoZapPlusCL antibiotics (Lonza). Next, cells were seeded in 96-well plates (100 µL of cells/ well) and cultivated at 37°C for 2 h
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using RNeasy mini kit (Qiagen) according to the manufacturer's instructions. Single-strand cDNA was prepared using the Ambion WT Expression kit
|
| Label |
Biotin
|
| Label protocol |
Probes were labeled using the GeneChip WT Terminal Labeling kit
|
| |
|
| Hybridization protocol |
5µg of fragmented cRNA was hybridized on a Affymetrix MoGene-1_0-st-v1. GeneChips were washed and stained in an Affymetrix Fluidics Station 450
|
| Scan protocol |
Scanning was performed on a GeneChip Scanner 3000 7G
|
| Data processing |
Data was processed with Affymetrix Expression Console 1.1 software using the RMA algorithm
|
| |
|
| Submission date |
Dec 20, 2012 |
| Last update date |
Apr 09, 2013 |
| Contact name |
Niels-Arne Münck |
| Organization name |
University of Münster
|
| Department |
Insitute of Immunology
|
| Street address |
Roentgenstraße 21
|
| City |
Münster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE43075 |
Inhibitor of NFκB ζ (IκBζ) is a transcriptional key regulator of monocyte chemoattractant protein-1 (MCP-1/CCL2) |
|
