Figure 6
From: Quantitative analysis of Hedgehog gradient formation using an inducible expression system
Non-Ptc containing Hh-GFP particles require cholesterol but not endocytosis. (A-C) Ptc co-localization with HhNp-GFP (A), HhN-GFP (B), and HhNp-GFP in the shits1 background (C) after expression induced for 8 hr. (A-C) Hh-GFP (green) labeled with Phalloidin (purple). (A'-C') Hh-GFP (green) labeled with Ptc (red). (A''-C'') Hh-GFP only. (A'''-C''') Ptc only. 4 classes of HhNp-GFP particles are seen: non-Phalloidin associated (cytoplasmic) with Ptc (white arrow), non-Phalloidin associated (cytoplasmic) without Ptc (white arrowhead), Phalloidin (membrane) associated with Ptc (yellow arrow), Phalloidin (membrane) associated without Ptc (yellow arrowhead). Most HhNp-GFP particles are membrane-associated and do not contain Ptc, but cytoplasmic particles have a relatively even distribution with and without Ptc. More HhN-GFP also localizes with Phalloidin, and almost all cytoplasmic HhN-GFP particles contain Ptc. HhNp-GFP particles in shits1 mutant background are Phalloidin-associated and many do not contain Ptc. The A/P boundary is marked by a solid white line. Scale bar: 5 μm.