Figure 9.

TX100/PBS-resistant cholesterol and TgGAP50 are completely extracted by mβCD. (A) The TX100/PBS-resistant fraction was incubated in PBS or in PBS containing 30 mM mβCD or 30 mM mβCD and 5 μg of cholesterol (CHOL) for 30 min at 37°C. TritonX-100 was added to 1%, and parasites were incubated on ice for 10 min. Soluble and insoluble fractions were separated by centrifugation at 14,000 × g for 10 min at 4°C and analyzed for the presence of TgGAP50 by SDS-PAGE and immunoblot analysis. (B) The TX100/PBS-resistant fraction of parasites prelabeled with BODIPY-CE was incubated in the presence or absence of 30 mM mβCD for 45 min at 37°C and analyzed by fluorescence microscopy. Bar, 2 μm.