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Status |
Public on Jun 29, 2017 |
Title |
siSLC25A10 [212_siSLC25A10_C07] |
Sample type |
RNA |
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Source name |
primary adipocyte culture, siSLC25A10
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Organism |
Homo sapiens |
Characteristics |
disease status: healthy, non-obese tissue: abdominal subcutaneous white adipose tissue (WAT) cell type: adipocytes sirna: siSLC25A10
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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.
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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.
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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.
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Label |
biotin
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Label protocol |
Total RNA were amplified and biotin labeled using the Whole Transcript (WT) Sense Target Labeling Protocol.
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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).
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Scan protocol |
Arrays were scanned using standardized Affymetrix protocols (Affymetrix Inc., Santa Clara, CA).
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Description |
212_siSLC25A10_C07
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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
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Submission date |
Feb 09, 2017 |
Last update date |
Jun 29, 2017 |
Contact name |
Ingrid Dahlman |
E-mail(s) |
[email protected]
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Organization name |
Karolinska
|
Street address |
Karolinska Huddinge
|
City |
Stockholm |
ZIP/Postal code |
14186 |
Country |
Sweden |
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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 |
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