Glucose Uptake in Adipocytes

Glucose uptake in primary adipocytes was determined as previously described (Gliemann et al., 1984 (link)). Briefly, cells (7.5% [vol/vol] suspension) were incubated with or without insulin (concentrations shown in Figure Legends) in KRBH buffer in triplicates for 30 min, followed by the addition of D-¹⁴C(U)-glucose (2.5 µl/ml, NEC042, Perkin Elmer, Waltham, USA), and an additional 30 min of incubation. The uptake was terminated by spinning 300 µl of each cell suspension in microtubes containing 80 µl dinonylphtalate oil. The cell fraction was collected, dissolved in scintillation fluid (Optima Gold, Perkin Elmer), and subjected to scintillation counting. Glucose uptake in 3T3-L1 adipocytes was performed as outlined (Roccisana et al., 2013 (link)).