Movie S2.

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Bidirectional transport of kinesin‐1 – DDB complexes by TIRF. Images at right are Qdot channel, showing kinesin‐DDB complexes moving slowly and bidirectionally along microtubules. Images at left are from GFP channel, showing streaming of excess free kinesin‐1 motors to the microtubule plus‐end; this information is used to assign microtubule polarity. Qdot and GFP movies were taken over different time windows. Arrows in first frame denote: cargos transported to the plus end (yellow arrow),cargos transported to the minus end (red arrow), and cargos that switch from plus‐end to minus‐end direction (white arrow).