Direct observation of Num1_CC-mediated minus end motility of dynein and dynactin. (A) Example kymographs of plus end–directed motility of dynein molecules along astral microtubules observed in uninduced GAL1p:num1_CC cells (left; –Num1_CC) or in cells overexpressing Num1_CC^LL/EE and Kip2 (right). (B and C) Example kymographs depicting minus end–directed motility of dynein or dynactin (i.e., Jnm1) along astral microtubules in cells overexpressing Num1_CC and either Kip2 (B) or Pac1 (C). Kymographs were generated from time-lapse images acquired using highly inclined and laminated optical sheet microscopy (see Materials and methods). Bars: (vertical) 1 min; (horizontal) 1 µm. (D) Velocity values for minus end–directed dynein runs observed in either Pac1- or Kip2-overexpressing cells. See also Fig. S5 and Videos 2 and 3.