|
| Status |
Public on Nov 21, 2007 |
| Title |
HUVEC_DMSO+TNF-α_6h_biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Confluent monolayers of HUVEC exposed to DMSO (final concentration in the culture medium <0.1%) for 45 min followed by addition of TNF-α (10 ng mL-1) for 6h.
|
| Organism |
Homo sapiens |
| Characteristics |
Normal human umbilical vein endothelial cells (HUVEC) under inflammatory conditions induced by cytokine TNF-α
|
| Treatment protocol |
HUVEC cells were treated as follows: i) direct treatment with dp3.9 apple procyanidins extract (final concentration in the culture medium, 10 μg mL-1) for 4h. As controls, cells were treated with DMSO (<0.1% in the culture medium) for 4h; ii) induction of inflammation with TNF-α: cells were treated with DMSO followed by TNF-α (10 ng mL-1) for 6 h. As controls, cells were treated with DMSO for 6h; iii) pre-co-treatment: cells were treated with dp3.9 (final concentration in the culture medium, 10 μg mL-1) for 45 min followed by treatment with TNF-α (10 ng/mL) for 6 h. As controls, cells were treated with DMSO for 45 min prior to TNF-a for 6 h. Treatments were all done in triplicate.
|
| Growth protocol |
Human umbilical vein endothelial cells (HUVEC) were obtained from Cambrex Bio Science (Wokingham, England), grown in EGM2 Bullet Kit (Lonza-Cambrex) and maintained at 37°C under a 5% CO2/95% air atmosphere at constant humidity. For the experiments, cells between passage number 2 and 4 (doubling population ≤ 10) were seeded onto 6-wells plates at a density of 3500 cells cm-2, and used on day 5 after seeding (confluent monolayers).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using an RNeasy ® Mini Kit (Qiagen) according to the manufacturer's instructions. RNA was eluted in RNAse free water, aliquoted and stored at –80ºC.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 ug total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome U133 plus 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
Gene expression data from unstimulated HUVEC exposed to DMSO and TNF-α for 6h
|
| Data processing |
The data were analyzed with Robust Multichip Average (RMA), using CEL files obtained from GCOS software (Affymetrix).
|
| |
|
| Submission date |
Nov 20, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
María-Teresa García-Conesa |
| E-mail(s) |
mtconesa@cebas.csic.es
|
| Phone |
+ 34 968 396276
|
| Organization name |
CEBAS-CSIC
|
| Department |
Quality, Safety and Bioactivity of Plant Foods
|
| Street address |
Campus de Espinardo
|
| City |
Espinardo |
| State/province |
Murcia |
| ZIP/Postal code |
30100 |
| Country |
Spain |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE9647 |
Comparative transcriptomic profiling of unstimulated and inflamed HUVEC exposed to apple procyanidin oligomers |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
