|
| Status |
Public on Apr 24, 2015 |
| Title |
Immature dendritic_cells_infected_HIV1+Vpx_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Immature dendritic cells,HIV1+Vpx infected, 72h, rep2
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Monocyte dendritic cells
treatment: 72h of HIV1+Vpx infection
|
| Treatment protocol |
Inmature and mature dendritic cells were infected with pseudotype HIV-1 (VSV-ΔenvGFP-200 ng of CA-p24) with or without Vpx for 72 h at 37ºC replacing the medium at 24h.
|
| Growth protocol |
Dendritic cells were diferentiated from monocyte cells extracted from peripheral blood from healthy donnors and treated with 20 ng/mL of IL-4 and GM-CSF during 7 days and growth with RPMI 1640 medium supplemented with 10% heat-inactivated fetal. Mature dendritic cells were differentiated with 20 ng/mL of LPS during 24h. The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using RNeasy Purification kit (QIAGEN, Valencia, CA) following the manufacturer's recommendations. The protocol includes on-column DNase digestion.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled anti-sense (aRNA) was prepared from 1.5 ug RNA using the One-Color Microarray-Based Gene Expression Analysis, Agilent Technologies according to the manufacturer's instructions. Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.65 ug of Cy3-labeled aRNA were hybridized for 17 hours at 65ºC in a Agilent hybridization oven (G2545A, Agilent Technologies) set to 10 rpm in a final concentration of 1x GEx Hybridization Buffer HI-RPM (Agilent Technologies). Arrays were washed and dried out using a centrifuge according to manufacturer's instructions.
|
| Scan protocol |
Arrays were scanned at 5μm resolution on an Agilent DNA Microarray Scanner (G2565BA, Agilent Technologies) using the default settings for 4x44k format one-color arrays.
|
| Description |
Gene expression after 72h of HIV1+Vpx infection
|
| Data processing |
Images provided by the scanner were analyzed using Feature Extraction software v10.7 (Agilent Technologies).
Notes : Entity List : Filtered on Flags [Detected, Not Detected]
Entities where atleast 1 out of 1 conditions have CV >= 1.0 percent
|
| |
|
| Submission date |
Apr 23, 2015 |
| Last update date |
Apr 24, 2015 |
| Contact name |
Esther Calonge |
| E-mail(s) |
ecalonge@isciii.es
|
| Organization name |
Instituto Salud Carlos III
|
| Department |
Centro Nacional Microbiologia
|
| Lab |
AIDS immunopathology
|
| Street address |
Ctra Pozuelo-Majadahonda Km 2
|
| City |
Madrid |
| ZIP/Postal code |
28220 |
| Country |
Spain |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE68191 |
Differential gene expression between infected and non-infected dendritic cells with HIV-1 |
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