 Competitive saturation binding assays were performed by incubating cerebellar
membrane protein extracts with 0.5-500 pM [125I]-labeled insulin, IGF-1, or
IGF-2 as tracer, in the presence or absence of 100 nM cold ligand. Membrane
bound tracer was precipitated and radioactivity in the supernatants (containing
free ligand) and pellets (containing bound ligand) was measured in a gamma
counter. Graphs depict specific binding (fmol/mg protein) ± S.D. relative to pM
input of radiolabeled (A-B) insulin, (C,D) IGF-1, and (E,F) IGF-2 in (A,C,E)
control (vehicle-treated) and (B,D,F) ethanol-exposed samples. The calculated
binding indices and inter-group statistical comparisons are provided in Table 1. |
