Overexpression of Num1_CC depletes dynein and dynactin from microtubule plus ends. (A) Schematic representation of Num1 and Num1_CC with domain structure indicated (CC1 and CC2, predicted coiled-coil domains; PH, plecktrin homology domain). (B) Diagram depicting experimental design (see text). (C) Western blot of GAL1p:num1_CC-13myc (wild-type or LL/EE mutant) cells grown in the absence or presence of galactose, as indicated, with loading control (anti–α-tubulin). (D) Representative images of GAL1p:num1_CC cells expressing mTurquoise2-Tub1 (α-tubulin) and either Dyn1-3mCherry (left) or Jnm1-3mCherry (right) used for quantitation in E and F. Cells were grown to mid-log phase in SD media supplemented with glucose (uninduced; −Num1_CC) or galactose plus raffinose (induced; +Num1_CC). 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. (E) The percentage of cells that exhibit plus end (red) or SPB (green) fluorescent foci is plotted for the strains shown in D. Plus end or SPB foci were identified in two-color movies and scored accordingly (see Materials and methods). Error bars represent the standard error of proportion (n ≥ 114 cells). (F) Box plot of fluorescence intensity values of plus end–associated Dyn1- or Jnm1-3mCherry foci (n ≥ 30 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 Video 1 and Figs. S1, S2, and S3.