Fig. 6

Effect of cholesterol depletion on CIR-dependent dissociation of pp125^Fak and pp59^Lyn from caveolin. Isolated rat adipocytes were incubated (see Materials and Methods) in the absence (Control) or presence of 10 µg/ml trypsin plus 0.5 M NaCl or 1.5 mM NEM. A portion of the trypsin/NaCl-treated adipocytes was reconstituted with trypsin/NaCl-extract. After further incubation (20 min, 37°C) in the absence or presence of increasing concentrations of PIG41, hcDIGs were prepared and used for immunoprecipitation (IP) of caveolin-1 in dissociating buffer. The immunoprecipitates were immunoblotted (IB) for pp125^Fak and pp59^Lyn. (A) Chemiluminescent images of a typical experiment repeated two times with similar results. (B) Quantitative evaluations of three different adipocyte incubations with measurements in triplicate each as percent of basal values (absence of PIG41) for control cells (mean ± SD) set at 100% in each case.