Figure 2

HhNp-GFP localizes at the membrane, in endocytic compartments, and extracellularly. (A-E) Localization of HhNp-GFP (green), expressed with Hh-Gal4, with FM4-64 (red, A-B), dextran (red, C-D), extracellular labeling with anti-GFP, and DCAD (red and blue, E). (B, D, E) Z-sections. HhNp-GFP (A') co-localizes with FM4-64 (A") at the membrane in the posterior (A/P boundary marked by the solid white line) as seen in the merge (A). Anterior HhNp-GFP appears in particles (arrows). Most of the particles localize apically (B). Many of the anterior HhNp-GFP particles co-localize (arrows in C-D) with dextran but some can be found without dextran (arrowheads in C-D). Incubation of anti-GFP in cold medium detects extracellular HhNp-GFP in the anterior apically in particles (arrows in E), and basolaterally both in particles (arrowheads in E) and with a membrane association (bracket in E). Scale bar: 8 μm