Figure 1

Purification of the adipocyte glycogen protoeme. Differentiated 3T3-L1 adipocytes were treated for 24 h with 2.5 mM glucose and 10 mM glucosamine, collected, freeze / thawed, sonicated and the 20 000 g supernatant (Ext) centrifuged at 400 000 g for 30 min. The glycogen pellet (P1) was resuspended and the process was repeated. The second glycogen pellet (P2) was resuspended in 50 mg malto-oligosaccharide (MO) / ml. The MO supernatant (SN3), or soluble fraction, was the reference preparation of glycogen-associated proteins used in this study. The MO pellet (P3), or insoluble fraction, was also trypsinized to provide a control comparative dataset to allow analysis of the stringency of the purification. The proteins illustrated above are a representative purification. Total amounts applied to the coomassie-G250 stained SDS-PAGE above, are 0.05% (Ext) or 2% (P1, P2, P3 and SN3) of the total sample.