|
| Status |
Public on Dec 15, 2008 |
| Title |
Macrophage Infected Day2 Rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Macrophages 2 days HIV-1 infected
|
| Organism |
Homo sapiens |
| Characteristics |
Primary human monocyte-derived macrophages
|
| Treatment protocol |
HIV-1JRFL was grown up in human peripheral blood mononuclear cells (PBMCs). Supernatant from HIV+ and HIV- PBMCs were applied to macrophages for 24 hours. Macrophages were rinsed with 1X PBS, and fresh media was applied. 50% media changes occurred every other day until RNA was extracted from cells.
|
| Growth protocol |
Primary human monocyte-derived macrophages were maintained in RPMI 1640 (Gibco), 10% FBS (Gibco), 50 ug/ml Penn/Strep (Gibco) at 37°C, 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from macrophages using Qiagen's RNeasy Kit according to manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
cRNA synthesis was performed with 10 ug of total cellular RNA, according to Affymetrix Expression Analysis Technical Manual. Double-stranded cDNA were synthesized according to a standardized protocol (SuperScript double-stranded cDNA synthesis kit; Invitrogen, Carlbad, CA). cRNA were transcribed in vitro, incorporating biotinylated nucleotides by using a BioArray high-yield RNA transcript labeling kit (T7) (Enzo Life Sciences, Farmingdale, NY).
|
| |
|
| Hybridization protocol |
Following fragmentation, Affymetrix human HG-U95Av2 oligonucleotide microarrays were hybridized for 16 hours at 45°C, stained with phycoerythrin-conjugated streptavidin and washed according to the Affymetrix protocol (EukGE-WS2v4) in an Affymetrix Fluidics Station.
|
| Scan protocol |
GeneChips were scanned with Affymetrix GeneChip 3000 scanner.
|
| Description |
none
|
| Data processing |
The arrays were analyzed with Microarray Suite version 4.0 (MAS 4.0) using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Oct 29, 2008 |
| Last update date |
Mar 16, 2009 |
| Contact name |
Maureen M. Goodenow |
| E-mail(s) |
goodenow@ufl.edu
|
| Phone |
352 273 8165
|
| Fax |
352 273 8184
|
| Organization name |
University of Florida
|
| Department |
Pathology, Immunology, & Laboratory Medicine
|
| Street address |
1376 Mowry Road
|
| City |
Gainesville |
| State/province |
FL |
| ZIP/Postal code |
32610-3633 |
| Country |
USA |
| |
|
| Platform ID |
GPL8300 |
| Series (1) |
| GSE13395 |
HIV-1 Activates Macrophages Independent of Toll-like Receptors |
|
