CD63-GFP traffics from secretory to endocytic compartments in SCs. (A–G) An 8-h pulse (at 28.5°C) of CD63-GFP expression was chased at 18°C for 0–60 h in virgin males, and proportions of cells with one or more LysoTracker red–positive iLEs (arrows in A, C, and E in apical sections; GFP positive in C and E) and mMVBLs (asterisks in B, D, and F in more basal views) that were CD63-GFP–positive were scored (G). Data shown are from a single representative experiment out of two repeats. The images in A–F are shown again in Fig. S3 alongside the corresponding single color channel images. Approximate outline of SC is marked in all images. **, P < 0.01; ***, P < 0.001; n > 24, pairwise comparisons, Fisher’s exact test. Bars, 5 µm.