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Status |
Public on Jun 03, 2016 |
Title |
HaCaT_ctl_3 |
Sample type |
RNA |
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|
Source name |
keratinocytes HaCaT, control transfection
|
Organism |
Homo sapiens |
Characteristics |
cell line: HaCaT transfection: control
|
Treatment protocol |
miR-21-3p mimic (Miridian, Thermo Scientific Dharmacon) or cel-miR-67-3p used as a control (Miridian, Thermo Scientific Dharmacon) were transfected to the cells at a concentration of 50 nM using the Lipofectamine RNAi max (Invitrogen). Cells were harvested 24h after transfection for gene expression analysis.
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Growth protocol |
Human immortalized keratinocytes HaCaT (Cell Line Service; Germany) cells were maintained in DMEM growth medium (Gibco) supplemented with 4500 mg/l glucose, 10% fetal bovine serum, 100 units/ml of penicillin G and 100 μg/ml of streptomycin. Cells were grown in a 5% CO2 atmosphere at 37 °C.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using TRIzol reagent (Invitrogen).
|
Label |
biotin
|
Label protocol |
For each sample, 100ng of total RNA were amplified using the Ambion® WT Expression Kit (4411973, life technologies) kit. 5.5μg of the cDNA was fragmented and labeled with GeneChip® WT Terminal Labeling kit (901525, Affymetrix).
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Hybridization protocol |
Affymetrix mouse gene 1.0ST arrays (Affymetrix, Santa Clara, CA, USA) were hybridized with 2.3μg of fragmented target, at 45°C for 16 hours washed and stained according to the protocol described in Affymetrix GeneChip® Expression Analysis Manual (Fluidics protocol FS450_0007).
|
Scan protocol |
The arrays were scanned with the GeneChip® Scanner 3000 7G (Affymetrix).
|
Description |
HaCat_ctrl_scr3
|
Data processing |
RMA algorithm was applied to data, using Affymetrix Expression Console software.
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Submission date |
Apr 19, 2016 |
Last update date |
Jun 03, 2016 |
Contact name |
Mark Ibberson |
Organization name |
SIB Swiss Institute of Bioinformatics
|
Department |
Vital-IT
|
Street address |
Genopode building
|
City |
Lausanne |
ZIP/Postal code |
CH-1015 |
Country |
Switzerland |
|
|
Platform ID |
GPL6244 |
Series (2) |
GSE80429 |
Identification of a novel PPARβ/δ / miR-21-3p axis in UV-induced skin inflammation [human mRNA] |
GSE80431 |
Identification of a novel PPARβ/δ / miR-21-3p axis in UV-induced skin inflammation |
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