Figure 4

Cholesterol restricts HhNp-GFP distribution but endocytosis is not required for distribution. (A-H) Induced expression of HhNp-GFP in wild-type (A-B) and shi^ts1 background (C-D) and HhN-GFP in wild-type (E-F) and shi^ts1 background (G-H). (A-H) 25 μm projections; (A'-H') 20 μm Z-section projections. At 8 hr, HhNp-GFP particles are found near the A/P boundary, marked by the solid white line (A, A'). After 24 hr, more particles can be found further away (B, B'). HhN-GFP particles are detected further from the A/P boundary than HhNp-GFP at both time points (E-F). When endocytosis is blocked, HhNp-GFP particles are still detected in anterior cells (C-D). In wild-type and shi^ts1 backgrounds, HhNp-GFP particles appear closer to the apical side (A'-D') as well as HhN-GFP in wild-type. When endocytosis is blocked, HhN-GFP moves into the anterior but there is reduced punctate staining and more membrane accumulation (G-H), primarily on the apical side of cells (G'-H'). Scale bar: 5 μm