|
| Status |
Public on Apr 09, 2014 |
| Title |
Vpr-Mac-Donor2 |
| Sample type |
RNA |
| |
|
| Source name |
Healthy Donor 2
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Monocyte Derived Macrophages (MDMs)
infection: HIV-1 Vpr
individual: healthy donor 2
|
| Treatment protocol |
MDMs were infected with adenovirus Vpr or ZsGreen1 at an MOI of 100. The cells were harvested for RNA extraction at 48 h post infection.
|
| Growth protocol |
Human PBMC were obtained using a standard Ficoll (Pharmacia ) gradient for heparinized blood extracted from two healthy individuals. CD14+ cells were isolated by positive selection using anti-human CD14+ magnetic beads (Miltenyi). Primary monocyte derived macrophages (MDMs) were generated by culturing CD14+ cells in RPMI (Sigma) medium supplemented with 10% FBS, 5% human AB serum, antibiotics and GlutaMax (Gibco) in the presence of recombinant human M-CSF (PeproTech).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
MDMs were washed three times with ice cold 1 x PBS, and total RNA was extracted using and RNeasy Mini Kit with DNase digestion, according to the manufacturer’s instructions (QIAgen).
|
| Label |
biotin
|
| Label protocol |
Biotin labeling of antisense cRNA was performed according to standard Affymetrix protocols
|
| |
|
| Hybridization protocol |
Sample hybridisation was performed for 16 hours according to standard Affymetrix protocols
|
| Scan protocol |
Samples were scanned using an Affymetrix GeneChip Scanner 3000 7G
|
| Data processing |
Affymetrix GeneChip Operating Software (GCOS) were used for processing scanning data. Normalization was performed using the RMA method.
|
| |
|
| Submission date |
Apr 08, 2014 |
| Last update date |
Apr 09, 2014 |
| Contact name |
Muhammad Atif Zahoor |
| Organization name |
University Health Network
|
| Street address |
101 College St.
|
| City |
Toronto |
| State/province |
Ontario |
| ZIP/Postal code |
M5G 1L7 |
| Country |
Canada |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE56591 |
Expression data from human monocyte derived macrophages infected with adenovirus expressing HIV-1 Vpr |
|
