Glucocorticoid target genes in 3T3-L1 adipocytes

Analysis of 3T3-L1 adipocytes treeated with dexamethasone (Dex) for 6 hours. Dex is a synthetic glucorticoid (GC) receptor agonist. Results provide insight into the effect of glucocorticoids on adipocytes. Differentiated 3T3-L1 adipocytes were cultured in DMEM supplemented with 5% stripped FBS for 24 hours and then treated with 500 nΜ Dex or an equal volume (0.05% v/v of media) of ethanol as a vehicle control for 6 hours at 37°C in DMEM with 190 5% stripped FBS. Total cellular RNA was isolated utilizing the NucleoSpin RNA II kit (Macherey-Nagel).RNA isolates were first quantified by standard spectrophotometry, and then qualitatively evaluated by capillary electrophoresis employing the Bio-Rad Experion system per the manufacturer’s instruction. The final labeled cRNA samples were hybridized overnight to Illumina mouseWG-6 BeadChip arrays, which was performed at UCSF Genomic Core. All treatments were done in triplicates and the same batch of microarrays were used for all treatments. The Illumina expression arrays were pre-processed using lumi package. The differential expression analysis was performed using the Limma package.