title: Illumina HiSeq 2500 paired end sequencing; RNA-seq comparison of primary mouse classical brown, beige, and white adipocytes
design: RNA-seq comparison of primary mouse classical brown, beige, and white adipocytes
library_construction_protocol: Primary stromal vascular (SV) cells were isolated from inguinal WAT or from interscapular BAT of the same C57BL/6 mice using Collagenase D (1.5 u/ml) and Dispase II (2.4 u/ml). To clearly define the molecular signatures of PPAR-gamma ligand-induced beige cells, we used C57BL/6 mice that possess less beige cells in the absence of environmental stimuli. SV cells were plated in collagen coated culture dishes in DMEM/F12 medium (D-glucose 17.51 mM) and differentiated as described previously (DOI: 10.1371/journal.pone.0049452). Beige cells were induced by a specific PPAR-gamma agonist rosiglitazone (1uM). RiboZol reagent (AMRESCO) following the manufacturer's protocol Ultralow DR library kit (NuGEN): www.nugeninc.com/nugen/index.cfm/products/cs/ngs/rna-seq-v2
attributes.tag: Experimental Factor: cell type
attributes.value: rosiglitazone-treated adipocyte (beige cell)