Dynein and dynactin localization during sliding events. (A) Time lapse images from a WT cell expressing Dyn1-3GFP and Jnm1-tdimer2 during a sliding event. Each image is a maximum intensity projection from a confocal Z series. Merged images show Dyn1-3GFP in green and Jnm1-tdimer2 in red. Arrowheads point to the microtubule plus end, and arrows point to stationary cortical foci. Dashed black lines are fiducial marks for assessing movement. Bars, 1 µm. (B) Localization of Dyn1-3GFP and Jnm1-tdimer2 at the plus end during the sliding event, from the boxed region in A. Bar, 1 µm. (C) Change in Dyn1-3GFP and Jnm1-tdimer2 signal during sliding events. Each dot represents the ratio of fluorescence intensity at t = 40 s of the sliding event divided by the intensity at t = 0 s. Black dots are measurements from the plus ends, and gray dots are measurements at stationary cortical foci. Red bars are the median. (D) Change in the ratio of Jnm1-tdimer2 to Dyn1-3GFP signal during sliding events. Each dot represents the fluorescence intensity of Jnm1-tdimer2 divided by the fluorescence intensity of Dyn1-3GFP at the same region. Black dots are measurements from the plus ends, and gray dots are measurements at stationary cortical foci. Red bars are the median. Cells were HU arrested, and 17 sliding events were analyzed. Asterisks denote a significant difference. *, P < 0.05; **, P < 0.005; ***, P < 0.0001; determined by t test.