2.4. Gene expression studies

RNA was extracted from differentiated OM adipocytes using Trizol following manufacturer's instructions. One microgram of RNA was used to synthetize cDNA and gene expression profiling was determined using RT2 Profiler human adipogenesis PCR arrays by assessing mRNA levels of 84 genes, including five "housekeeping genes" according to manufacturer's protocol. The list of genes included Cyclin D1 (CCND1), Fatty Acid Binding Protein 4 (FABP4), TSC22 Domain Family Member 3 (TSC22D3) and Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1 Beta (PPARGC1B, also known as PGC1beta). Data were normalized with the internal housekeeping genes and ΔΔCt was calculated using ΔCt from IS as the control group according to manufacturer's protocol.