Late endosomal DRMs are sensitive to cholesterol affecting reagents.

<p>Late endosomes were prepared from BHK cells using a sucrose step gradient, treated or not with either filipin (1μg/ml for 1 h at 37°C) or saponin (0.4% for 1 h at 4°C) and then submitted to solubilization in 1% Triton X-100 at 4°C. The lysat was submitted to an OptiPrep flotation gradient and 6 fractions of 400 µl were collected. Each fraction was analyzed by SDS-PAGE followed by an aerolysin overlay to identify the GPI-anchored proteins of BKH cells: N-CAM-120, semaphorin-7 (Sema-7), CD14 and Thy-1.