3T3L1 adipocytes were plated at 50,000 cells/well/100 µl of DMEM/FBS in 96-well plate for 5 hours, then serum deprived for 1 hour with or without leptin (100 nM). Insulin at 10 nM was then added into the well and incubated for a further 30 minutes at 37°C, 5% CO2 incubator. At the end of the incubation time, 100 µl of fatty acid was added into the well, and kinetic readings were started immediately with a FlexStation® 3 Reader. Data courtesy of Cambridge Biotechnology. A: insulin, B: insulin and leptin, C: basal.