|
| Status |
Public on Dec 19, 2016 |
| Title |
Control hMADS cells, replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
hMADS cells, D18, untreated
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: hMADS cells
time of differentiation: Day 18
cell type: Adipocyte
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRI-Reagent kit (Euromedex, Souffelweyersheim, France) according to the manufacturer’s instructions.
|
| Label |
Cy5
|
| Label protocol |
Total RNA quality was checked by capillary electrophoresis (Experion, BioRad) and quantified using Nanodrop ND-1000 Spectrophotometer (Labtech). Only good quality RNA extracts were used for microarray hybridization. Fluorescent probes were obtained by amplifying and labelling 500ng of RNA with the Quick Amp Labeling kit, two-color (Agilent).
|
| |
|
| Channel 2 |
| Source name |
Reference
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: hMADS cells
time of differentiation: Day 9
cell type: Adipocyte
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRI-Reagent kit (Euromedex, Souffelweyersheim, France) according to the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Total RNA quality was checked by capillary electrophoresis (Experion, BioRad) and quantified using Nanodrop ND-1000 Spectrophotometer (Labtech). Only good quality RNA extracts were used for microarray hybridization. Fluorescent probes were obtained by amplifying and labelling 500ng of RNA with the Quick Amp Labeling kit, two-color (Agilent).
|
| |
|
| |
| Hybridization protocol |
825 ng of Cy5 labelled probe was cohybridized with the same amount of Cy3 labelled control probe. We used the Whole Human Genome Microarray Kit, 4x44K from Agilent with the gene expression hybridization kit according to the manufacturer’s protocol.
|
| Scan protocol |
Micro-arrays were scanned on a InnoScan 710A scanner (Innopsys, Carbonne, France, EU) and data were extracted with Mapix software version 5.1.1 (Innopsys).
|
| Data processing |
Labeling and hybridization qualities were verified with Agilent’s spike in kit. Features with a signal to background ratio strictly below 1.2 in both color as well as control spots were discarded. The data without background substraction were normalized with the Lowess procedure and quantiles method by use of R.
|
| |
|
| Submission date |
Jul 24, 2015 |
| Last update date |
Dec 19, 2016 |
| Contact name |
Nathalie Viguerie |
| E-mail(s) |
[email protected]
|
| Phone |
+33 5 61 32 56 31
|
| Organization name |
INSERM-Université Paul Sabatier
|
| Department |
Institute of Metabolic and Cardiovascular Diseases, UMR1048
|
| Lab |
Obesity research laboratory, CHU Rangueil, bâtiment L4
|
| Street address |
1 avenue Jean Poulhès BP 84225
|
| City |
Toulouse cedex 4 |
| ZIP/Postal code |
31432 |
| Country |
France |
| |
|
| Platform ID |
GPL13497 |
| Series (1) |
| GSE71293 |
Transcriptional profile of human white adipocytes (hMADS cells) after white-to-brown conversion |
|
