Figure 2.

Concentration-dependence of wortmannin-induced redistribution of EEA1. 3T3-L1 adipocytes were treated with increasing concentrations of wortmannin (WTM) for 15 min, and fractionated into a crude membrane (P) and cytosolic (C) fraction. Proportional amounts of the two fractions were separated by SDS/PAGE and analyzed by immunoblotting with human anti-EEA1 antiserum and ¹²⁵I-labeled protein A. A single protein band at 170 kDa was detected (Inset) in both cytosol (C) and pelleted membrane (P) fractions. The intensity of the bands was quantified by densitometric scanning. Plotted are the ratios (P/C) of the densitometric values obtained.