Dyn1_HL3 is less susceptible to Num1_CC-mediated plus end depletion. (A) Diagram depicting the Dyn1_HL3 high Pac1 affinity mutant, in which a helical linker has been inserted between the dynein tail and motor domains (Markus and Lee, 2011). (B) Representative images of GAL1p:num1_CC cells expressing mTurquoise2-Tub1 and either Dyn1_HL3-3YFP (left) or Dyn1_HL3 and Pac1-3mCherry (right) used for quantitation in C and D. Each image is a maximum-intensity projection of a 2-µm Z-stack of wide-field images. Arrows indicate plus end foci, and arrowheads indicate SPB foci. Bars, 2 µm. (C) The percentage of cells that exhibit plus end (red) or SPB (green) fluorescent Dyn1_HL3-3YFP (left) or Pac1-3mCherry (right) foci is plotted for the cells shown in B. Error bars represent the standard error of proportion (n ≥ 119 cells). (D) Box plot of fluorescence intensity values of plus end–associated Dyn1_HL3-3YFP or Pac1-3mCherry (n ≥ 33 foci). Whiskers define the range of data, boxes encompass the 25th to 75th quartiles, the line depicts the median value, and the “x” depicts the mean value. See also Fig. S5.