|
| Status |
Public on Nov 01, 2013 |
| Title |
Donor B Untr |
| Sample type |
RNA |
| |
|
| Source name |
Mono-DC, Untreated
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: monocyte-derived Dendritic cells
subject: donor B
treatment: Untreated
|
| Treatment protocol |
Human DC derived from monocytes from three different healthy volunteers were left untreated or stimulated for 4 and 12 h with beta-glucan, in absence or presence of IL-1RA, in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum. The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2.
|
| Growth protocol |
Human monocyte-derived DC were obtained by a 6/7-d cultures of freshly isolated monocytes in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum plus recombinant human IL-4 (10 ng/ml) and GM-CSF (50 ng/ml). The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the Rneasy Mini Kit (QIAGEN) and residual genomic DNA was digested twice. RNA was quantified using a NanoDrop-1000 spectrophotometer and its quality was assessed with an Agilent Bioanalyzer.
|
| Label |
Cy3
|
| Label protocol |
First and second strand cDNA was prepared from the total RNA samples. cRNA target was prepared from the DNA template and verified on the Bioanalyzer. cRNA was fragmented to uniform size and verified on the Bioanalyzer.
|
| |
|
| Hybridization protocol |
cRNA was fragmented to uniform size and hybridized to Agilent Whole Genome 4x44K arrays. Arrays were hybridized at 65°C for 17 h in a rotating incubator, and washed afterwards at 37°C for 1 min.
|
| Scan protocol |
Slides were washed and scanned on an Agilent G2565 Microarray Scanner at 5 micron resolution
|
| Description |
Gene expression analysis of untreated mono-DCs
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) and Partek Genomics Suite 6.6
|
| |
|
| Submission date |
Nov 09, 2012 |
| Last update date |
Nov 01, 2013 |
| Contact name |
Marco Cardone |
| E-mail(s) |
cardonem@mail.nih.gov
|
| Phone |
3012284387
|
| Organization name |
NCI/NIH
|
| Department |
CIP
|
| Street address |
1050 Boyles Street
|
| City |
Frederick |
| State/province |
MD |
| ZIP/Postal code |
21702 |
| Country |
USA |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE42189 |
The Cytokines Interleukin-1 and Interferon-gamma Differentially Program Beta-glucan–activated Dendritic Cells via IkappaB-zeta Modulation |
|
