|
| Status |
Public on Jan 30, 2022 |
| Title |
HepaRG cells treated with sVE and OA [sVE_OA 3] |
| Sample type |
RNA |
| |
|
| Source name |
Treated with William E complete medium with oleic acid [200 µM]+ synthetic all-rac-α-Tocopherol [25nM]
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: hepatic cell line
cell type: Immortalized hepatic cell line terminally differentiated
age: Adult
treatment: treated with sVE and OA
|
| Treatment protocol |
At 70% confluence, HepaRG cells were incubated for 24 hrs with either the vehicle or the three different sources of Vitamin E at final concentration of α-Tocopherol in the colture medium of 25 nM. Subsequently HepaRG were incubated with OA at the final concentration in the colture medium of 200 µM. Cell treatments with this OA lasted for 48 hrs.
|
| Growth protocol |
Cells were seeded at a density of 10,000 cells/well and kept at 37˚C in a humidified 5% CO2 cell culture incubator.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells lysates using an QIAsymphony RNA Kit (Qiagen,) with the QIAsymphony SP instrument (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the standard ThermoFisher Scientific protocol from 6 ug total RNA (GeneChip™ 3’ IVT Pico Kit Manual Workflow).
|
| |
|
| Hybridization protocol |
Fragmented and labeled cDNA was prepared according to the manufacturer's instructions and hybridized to a Human Clariom Go Screen 384-array plate for 17 h at 45°C on a GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific). Arrays were washed and stained on a GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific).
|
| Scan protocol |
Arrays were scanned using the GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific). Affymetrix GeneChip Command Console Software (AGCC, ThermoFisher Scientific) was used for GeneTitan MC Instrument control.
|
| Data processing |
Data analysis were performed using Transcriptomic Analysis Console Software (TAC, ThermoFisher Scientific) that provides quality control analysis, performs normalization and summarization, based on the Signal Space Transformation-Robust Multi-Chip Analysis (SST-RMA) analysis algorithm, and provides a list of differentially expressed genes (Limma Bioconductor package, pvalue<0,05) from the biological duplicates for WGO-VE_OA or triplicates of the other condition.
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| |
|
| Submission date |
Jan 28, 2022 |
| Last update date |
Jan 30, 2022 |
| Contact name |
Jacopo Lucci |
| E-mail(s) |
jlucci@biostherapy.it
|
| Phone |
+393428018168
|
| Organization name |
Bios-Therapy, Physiological Systems For Health S.p.A
|
| Lab |
Laboratory of system biology
|
| Street address |
Loc. Aboca, 20
|
| City |
Sansepolcro |
| State/province |
Arezzo |
| ZIP/Postal code |
52037 |
| Country |
Italy |
| |
|
| Platform ID |
GPL31262 |
| Series (1) |
| GSE195619 |
Expression data from HepaRG human progenitor hepatic cells treated with different sources of Vitamin E and wheat germ oil. |
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