Fig. 3

Effect of cholesterol depletion on PIG41- and glimepiride-induced redistribution and activation of signaling components. Isolated rat adipocytes were incubated in the absence or presence of 10 mM mβCD (50 min, 30°C) and subsequently without (Con) or with 10 nM insulin (Ins), 3 µM PIG41, or 30 µM glimepiride (Am) (15 min, 37°C). hcDIGs were prepared and used for immunoprecipitation (IP) of caveolin-1 in nondissociating buffer. The immunoprecipitates were immunoblotted (IB) for pp125^Fak, pp59^Lyn, and caveolin-1, and photoaffinity labeled for Gce1 (PL). From total cell lysates, pp59^Lyn, IRS-1, and IRβ were immunoprecipitated and then immunoblotted for phosphotyrosine (pY). Representative phosphor/chemiluminescent images repeated three times with similar results are shown.