Fig. 2.

HTT moves within discrete Rab4-containing vesicles, which also contain Syt, Syb and Rab11. a Representative kymographs (x-axis = distance (μm) || y-axis = time(s)) from simultaneous dual-color movies from larvae expressing both HTT25Q-eGFP (green) and Rab4-mRFP (red). Co-migrating tracks that contain HTT and Rab4 are seen (yellow arrow). Note that green or red only tracks are also seen (arrowheads). Representative kymographs from larvae expressing HTT15Q-mRFP (red) and either Syt-GFP, nSyb-GFP, APP-YFP or Rab11-GFP (green). Yellow trajectories show overlaid red and green tracks. Scale = 5 μm. b Representative kymographs from larvae expressing Rab4-mRFP (red) and either Syt-GFP, nSyb-GFP, APP-YFP, YFP-Rab3 or Rab11-GFP (green). Yellow trajectories show overlaid red and green tracks. Scale = 5 μm. c Quantification of the percentage (%) of trajectories that associate with HTT trajectories (either GFP or RFP) show that ~ 21% of Rab4-mRFP trajectories overlay with HTT25Q-eGFP tracks while ~ 8% of APP-YFP trajectories overlay with HTT15Q-mRFP tracks. n = 5. d Quantification of the percentage (%) of trajectories that associate with Rab4-mRFP trajectories show that ~ 33% of Syt-GFP trajectories overlay with Rab4-mRFP tracks while 0% of APP-YFP trajectories overlay with Rab4mRFP tracks. n = 5. e Proportional Venn diagram (generated using OriginLab & RStudio) display the percent of trajectories that associate between either Rab4, HTT, nSyb, Syt or APP trajectories relative to the total number of trajectories quantified (top). An additional proportional Venn diagram displays the percent of trajectories that associate between either Rab4, HTT, or Rab11 (bottom). f Co-localization pixel-heat maps (ImageJ.Coloc2) generated for each of the representative movies from panel A and panel B. x-axis = GFP/YFP channel. Y-axis = RFP channel. A line near 45 degrees with a higher Pearson’s correlation coefficients R-value near 1.0 corresponds to co-localization