Fig. 3

Kinesin-1, dynein, and hip1 aid the motility of HTT-Rab4 in axons. a Representative movies with kymographs from Rab4-mRFP larvae alone or in the context of 50% reduction of klc^8ex94−/+ or robl^K−/+. Bar = 10 μm. b Quantification of duration weighted segmental velocity (μm/s) indicates significant decreases in the anterograde Rab4-mRFP velocities with 50% reduction of kinesin (p < 0.001) or dynein (p < 0.01). 50% reduction of dynein significantly decrease retrograde Rab4-mRFP velocities (p < 0.001), 50% reduction of kinesin did not (ns). n = 10. > 500 vesicles. c Sub-pixel high-resolution immunofluorescence analysis show Rab4-mRFP (red arrows) and klc (green arrows) co-localize (yellow arrows) within larval segmental nerves. Intensity-pixel plots show overlapping (purple arrows) peak intensities for both Rab4 (red) and klc (green). Rab4-mRFP (red arrows), HTT25Q-eGFP (green arrows) and DIC (blue arrows) also co-localize (white arrows). Intensity-pixel plots show overlapping (purple arrows) peak intensities for Rab4 (red) HTT (green) and DIC (blue). X axis = distance (μm), Y axis = intensity (au). Bar = 1 μm. n = 5. d, e Representative images of Rab4-mRFP or HTT15QmRFP larval nerves alone or in the context of 50% reduction of hip1, rip11, milt, nuf or nmo. Bar = 10 μm. f Quantification of the number (#) of Rab4 blocks per larvae reveal that larvae expressing Rab4-mRFP in the context of 50% reduction of rip11 (p < 0.01) or hip1 (p < 0.01) show Rab4-mRFP blocks. n = 5. g Quantification of the number (#) of blocks per larvae reveal that larvae expressing HTT15Q-mRFP in the context of 50% reduction of hip1 (p < 0.01) show HTT15Q-mRFP blocks. n = 5. Statistical significance was determined using the two-sample two-tailed Student’s t-test. h Representative movies with the corresponding kymographs from larvae expressing Rab4-mRFP alone or in the context of 50% reduction of hip1. Quantification of duration weighted segmental velocity (μm/s) indicates significant decreases in anterograde Rab4-mRFP velocities with 50% reduction of hip1 (p < 0.001) and in retrograde Rab4-mRFP velocities (p < 0.001). n = 10, > 500 vesicles. i Representative movies with the corresponding kymographs from larvae expressing HTT15Q-mRFP alone or in the context of 50% reduction of hip1. Quantification of duration weighted segmental velocity (μm/s) indicates significant decreases in anterograde velocities with 50% reduction of hip1 (p < 0.01) and significant decreases in retrograde HTT15Q-mRFP velocities (p < 0.01). n = 10. > 500 vesicles. Statistical significance was determined using the two-sample two-tailed Student’s t-test. ns = p > 0.01, *p < 0.01, **p < 0.001