FIGURE 6.

The binding of NPC1L1 to Rab11a and myosin Vb is regulated by cholesterol depletion. A, plasmids encoding NPC1L1-EGFP and Rab11a-T7 were co-transfected into CRL-1601 cells. 48 h later, the cells were depleted of cholesterol by treatment with 1% CDX for various time durations. Then the cells were harvested, and immunoprecipitation (IP) was performed by pulling down EGFP-tagged NPC1L1 with anti-EGFP antibody-coupled agarose. Immunoblot (IB) analysis was carried out using the indicated antibodies. B, plasmids encoding NPC1L1-T7 and EGFP-myosin Vb were co-transfected into CRL-1601 cells. 48 h later, the cells were depleted of cholesterol by treatment 1% CDX for various durations of time. Then the cells were harvested, and immunoprecipitation was performed by pulling down EGFP-tagged myosin Vb with anti-EGFP antibody-coupled agarose. Immunoblot analysis was carried out using the indicated antibodies. Results shown are representative of three independent experiments.