Visceral fat biopsies were obtained intra-operatively from the omentum. Samples were rinsed with PBS to remove excess blood, minced, immediately frozen in liquid nitrogen and stored at −80°C.
Growth protocol
NA
Extracted molecule
total RNA
Extraction protocol
Total RNA was obtained using RNeasy Lipid Tissue Mini Kit with DNase treatment (Qiagen, Valencia, CA) according to the manufacturer's protocol. The integrity of purified total RNA samples was assessed qualitatively on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
Label
Biotin labeled cRNA
Label protocol
100 ng of total RNA was labeled using Affymetrix GeneChip Whole Transcript (WT) Sense Target Labeling Assay according to the manufacturer's protocols.
Hybridization protocol
Standard hybridization protocol was used as recommended by Affymterix
Scan protocol
Microarrays were scanned using Affymetrix GeneChip Scanner, and the GeneChip Operating Software (GCOS) was used for image analysis.
Data processing
Background adjustment and quantile normalization using the Robust Multichip Average algorithm using Log2 transformed intensities.
