Endogenous localization of kinesin-1/UNC-116. (A) Labeling scheme of endogenous kinesin-1/UNC-116. (B) Representative images showing the localization of kinesin-1::3xSplitGFP (UNC-116::3xSplitGFP) in the axon and the dendrite. + indicates the DA9 dendrite, whereas — indicates the VA12 axon. Note that the dendritic kinesin-1::3xSplitGFP is much less abundant. Scale bars = 5 μm. (C) Quantification of kinesin-1::3xSplitGFP intensity in DA9 axon and dendrite. Each data point represents one animal. Number of animals: 8 (axon) and 10 (dendrite). **P < 0.01 (t test). (D) Kymograph showing anterograde and retrograde trafficking events of kinesin-1::3xSplitGFP in the axon with (left) or without (right) manual traces. Green: anterograde; salmon: retrograde. (E–G) Quantification of anterograde and retrograde trafficking events of kinesin-1::3xSplitGFP in the axon. (E) Percentage of anterograde and retrograde trafficking events, respectively. Error bars represent 95% confidence intervals. (F and G) Speeds and run lengths of anterograde and retrograde trafficking events, respectively. Violin plots with median and quartiles are shown. Total number of events analyzed: anterograde = 339; retrograde = 271. Number of animals: 12. (H) Representative images showing that kinesin-1::3xSplitGFP is enriched at the leading end of a mitochondrion moving anterogradely. Arrow heads highlight the accumulation of kinesin-1::3xSplitGFP. (I) Normalized intensity of kinesin-1::3xSplitGFP along mitochondria length averaged across different timepoints during anterograde trafficking, and then averaged across seven different mitochondria from seven different animals. Leading end is on the left and trailing end on the right. Error bars represent SEM. Data are pooled from four independent experiments.