|
| Status |
Public on Aug 21, 2011 |
| Title |
Epidydimal white adipose tissue from ASC-/- after 16 weeks of high fat diet feeding, replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Adipose tissue from male ASC-/- animal (C57/Bl6 background)
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57/Bl6
genotype/variation: ASC1 null
gender: male
tissue: epididymal white adipose tissue
|
| Treatment protocol |
White adipose tissue was dissected from all 3 genotypes after 16 weeks of high fat diet-feeding. Adipose tissue RNA samples from 3-4 mice per experimental group were used for microarray analysis.
|
| Growth protocol |
Male wild-type (control) (C57/BL6 background) and ASC-/- and Casp1-/- mice (C57/BL6 background), aged 8-12 weeks at the start of the intervention, were maintained on a high fat diet (45 energy% of fat) for 16 weeks. After the diet intervention, mice were killed and the adipose tissue was removed.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from mouse adipose tissue using TRIzol reagent and whereafter purified total RNA was isolated using Qiagen RNEasy columns. RNA integrity was checked on chip analysis (Agilent 2100 bioanalyzer, Agilent Technologies, Amsterdam, the Netherlands) according to the manufacturer's instructions. RNA was judged as suitable for array hybridization only if samples exhibited intact bands corresponding to the 18S and 28S ribosomal RNA subunits, and displayed no chromosomal peaks or RNA degradation products (RNA Integrity Number > 8.0).
|
| Label |
biotin
|
| Label protocol |
All target labeling reagents were purchased from Affymetrix (Santa Clara, CA). Double-stranded cDNAs were synthesized from 1ug total RNA through reverse transcription with an oligo-dT primer containing the T7 RNA polymerase promoter and double strand conversion using the cDNA Synthesis System. Biotin-labeled cRNA was generated from the cDNA and used to probe Affymetrix Mouse Genome HT_MG-430 PM arrays.
|
| |
|
| Hybridization protocol |
The GeneChip HT MG-430 PM 24 Array Plate consists of 24 single MG-430 PM arrays arranged into standard 96 well plate format. Array hybridization, washing and scanning were performed according to the manufacturer’s recommendations on a GeneTitan Instrument.
|
| Scan protocol |
Arrays were scanned on an Affymetrix GeneTitan instrument.
|
| Data processing |
Expression estimates were calculated using RMA in Bioconductor.
|
| |
|
| Submission date |
Nov 08, 2010 |
| Last update date |
Aug 21, 2011 |
| Contact name |
Guido Hooiveld |
| E-mail(s) |
guido.hooiveld@wur.nl
|
| Organization name |
Wageningen University
|
| Department |
Div. Human Nutrition & Health
|
| Lab |
Nutrition, Metabolism & Genomics Group
|
| Street address |
HELIX, Stippeneng 4
|
| City |
Wageningen |
| ZIP/Postal code |
NL-6708WE |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL11180 |
| Series (1) |
| GSE25205 |
The inflammasome is a central player in the induction of obesity and insulin resistance |
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