|
| Status |
Public on Feb 13, 2021 |
| Title |
BMDM_2_PBS_1h |
| Sample type |
RNA |
| |
|
| Source name |
Primary mouse bone marrow-derived macrophages
|
| Organism |
Mus musculus |
| Characteristics |
cell type: BMDM
agent: PBS
time: 1h
|
| Treatment protocol |
Hepcidin (Peptide International) was added to the wells at 10µg/ml. The BMDMs were incubated for one hour and lysed with Trizol.
|
| Growth protocol |
Bone marrow-derived macrophages (BMDMs) were obtained from bone marrow (BM) precursor cells. In brief, total BM was obtained from 8-10 week old WT by flushing femur and tibiae BM with Dulbecco’s modified Eagle’s medium (DMEM). Cells were cultured in plastic tissue culture dishes (150 mm) in 25 ml of DMEM containing 20% FBS (Fetal Bovine Serum) and 30% L cell-conditioned medium as a source of M-CSF. They were then incubated at 37°C in a humidified 5% CO2 atmosphere. After 7 days of culture, a homogeneous population of adherent BMDMs was obtained. BMDMs were scrapped and seeded at 150 000 cells per well into 24-well plate in 0,5 ml of DMEM containing 15% L cell-conditioned medium and 10% FBS. The following day, we change media to DMEM alone for 1 hour and hepcidin (Peptide International) was added to the wells at 1 µg/ml. The BMDMs were incubated for 1 and 3 hours and lysed with Trizol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix protocol from total RNA (Following the GeneChip whole transcript (WT) Sense Target labelling assay kit Affymetrix)
|
| |
|
| Hybridization protocol |
Following fragmentation, cDNA were hybridized for 16 hr at 45C on GeneChip mouse Gene 2.0 from Affymetrix GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GCS 3000 scanner (Affymetrix)
|
| Description |
02-17
|
| Data processing |
Data were normalized using RMA and statistical analysis was realized on Partek Genomics Suite. CDF file V21 was used for annotation.
|
| |
|
| Submission date |
Feb 21, 2017 |
| Last update date |
Feb 13, 2021 |
| Contact name |
Franck Letourneur |
| E-mail(s) |
franck.letourneur@inserm.fr
|
| Organization name |
INSERM
|
| Department |
U1016
|
| Lab |
genomic
|
| Street address |
22 rue mechain
|
| City |
Paris |
| ZIP/Postal code |
75014 |
| Country |
France |
| |
|
| Platform ID |
GPL23092 |
| Series (1) |
| GSE95162 |
Hepcidin: a key regulator of inflammation |
|
