|
| Status |
Public on Jun 29, 2017 |
| Title |
siNEGC [184_siRNANegC_C05] |
| Sample type |
RNA |
| |
|
| Source name |
primary adipocyte culture, siNEGC
|
| Organism |
Homo sapiens |
| Characteristics |
disease status: healthy, non-obese
tissue: abdominal subcutaneous white adipose tissue (WAT)
cell type: adipocytes
sirna: siNEGC
|
| Treatment protocol |
In vitro differentiated adipocytes (day 10-12 post induction) were transfected with ON-TARGETplus SMARTpool small interfering RNAs (siRNAs) targeting KLF15, SLC25A10 or non-targeting siRNA pool at 40 nM final concentration (Dharmacon, Lafayette, CO) in 24- or 48-well plates and HiPerfect Transfection Reagent (Qiagen), respectively 4.5 or 2 µl according to the manufacturer's protocol. The cells were incubated for 48 or 72 hours at which time in vitro lipogenesis was assessed in cells plated on 48-well plates and RNA were collected.
|
| Growth protocol |
Primary adipocyte culture for in vitro studies was obtained from subcutaneous WAT from healthy non-obese men and women (BMI <30 kg/m2) undergoing cosmetic liposuction. The stroma vascular fraction (SVF) cells were isolated. The precursor cells obtained from separate individuals were not mixed. Part of plastic-adherent SVF cells were directly plated (30,000-50,000 cells/cm2) and differentiated to adipocytes as described. The degree of differentiation was controlled under the microscope as accumulation of lipids, and response to insulin in lipogenesis assay was evaluated.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using NucleoSpin RNA II kit (Macherey-Nagel, Düren, Germany) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Total RNA were amplified and biotin labeled using the Whole Transcript (WT) Sense Target Labeling Protocol.
|
| |
|
| Hybridization protocol |
From non-degraded high-quality total RNA we prepared and hybridized biotinylated complementary RNA to Gene 2.1 ST Arrays, and then washed and stained slides using standardized protocols (Affymetrix Inc., Santa Clara, CA).
|
| Scan protocol |
Arrays were scanned using standardized Affymetrix protocols (Affymetrix Inc., Santa Clara, CA).
|
| Description |
184_siRNANegC_C05
|
| Data processing |
These arrays were pre-processed with RMA, which includes normalization, summarization of probes to probesets, and background correction.
Analysis Method: rma-gene-full
Analysis Software: Expression Console
Data Transformation: log2
Annotations: hg19/GRCh37
NetAffx build: 33
|
| |
|
| Submission date |
Feb 09, 2017 |
| Last update date |
Jun 29, 2017 |
| Contact name |
Ingrid Dahlman |
| E-mail(s) |
[email protected]
|
| Organization name |
Karolinska
|
| Street address |
Karolinska Huddinge
|
| City |
Stockholm |
| ZIP/Postal code |
14186 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL17692 |
| Series (2) |
| GSE94751 |
Global transcriptome profiling identifies KLF15 and SLC25A10 as regulators of adipocytes insulin sensitivity in obese women [siRNA] |
| GSE94753 |
Global transcriptome profiling identifies KLF15 and SLC25A10 as regulators of adipocytes insulin sensitivity in obese women |
|
