|
| Status |
Public on Sep 01, 2013 |
| Title |
Control-biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Control-biological replicate 3
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: Control
cell line: HeLa
|
| Treatment protocol |
Purified T3s were added to the culture medium for the indicated period of time. Final TRF concentration in culture media was set at 10 or 20 µg/ml, to standardize the present study with our previous reports. Control cells were treated with the same volumes of DMSO and/or ethanol vehicle alone.
|
| Growth protocol |
HeLa cells were obtained from the American Tissue Culture Collection (Manassas, VA, USA). Cells were grown in DMEM medium (Euroclone) supplemented with 10% Foetal Bovine Serum (FBS, Sigma-Aldrich), Pen/Strep (Euroclone), 2mM glutamine (Euroclone) and non-essential aminoacid (Sigma Aldrich). Before any experimental session, cells were synchronized in G1/G0 by starvation in serum-free medium for two days. Once synchronized, 300.000 cells were seeded onto multi well plates or phenol red DMEM according to experimental planning, where appropriate, incubated with ICI 182.780 (10-5 M in ethanol) for 30 minute
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Protocol of DNA extraction was adapted from Gooch and Yee (Gooch, 1999)with some modification described in Comitato et al., 2010
|
| Label |
biotin
|
| Label protocol |
2μg of high quality total RNA was used with the Affymetrix Eukaryotic One-Cycle Target Labeling and Control reagents to generate biotin-labeled antisense cRNA
|
| |
|
| Hybridization protocol |
The labeled cRNA was hybridized to the NuGO Affymetrix Human Genechip NuGO_Hs1a520180 (custom designed by the European Nutrigenomics Organisation NuGO, consisting of 23,941
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Geneexpression data from HeLa cells untreated
|
| Data processing |
Three biological replicates were generated for each experimental condition. Microarrays statistical analysis was performed using oneChannelGUI R package, Raw signal intensities were normalized using GCRMA method as background correction
|
| |
|
| Submission date |
Jul 10, 2013 |
| Last update date |
Sep 01, 2013 |
| Contact name |
Guido Leoni |
| E-mail(s) |
[email protected]
|
| Organization name |
Sapienza
|
| Department |
Physics
|
| Street address |
P.le Aldo Moro n5
|
| City |
Rome |
| ZIP/Postal code |
00100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL7020 |
| Series (1) |
| GSE48668 |
Expression data from HeLa cells treated with tocotrienols |
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