Dynein heavy chain asymmetry is lost in DYNLL1-, CHICA-, and HMMR-depleted cells. (A) GFP-dynein heavy chain (DYNC1H1) cells were transfected with control, CHICA, HMMR, DYNLL1, and HURP siRNA duplexes for 72 h. The cells were then imaged using an Ultraview Vox spinning-disk confocal system. Images of metaphase cells were taken from a single z-plane at one time point. Exposure times were 100 msec for GFP-DYNC1H1 using 50% laser power. Graphs representing the intensity profiles were generated with NIH ImageJ. A profile plot was made from a selection of 20 pixels width crossing the mitotic spindle at both spindle poles. Intensity values were extracted. An average background value was subtracted and values were normalized to the brightest point corresponding to 100%. Two representative example images and associated graphs are shown. Bar, 10 µm. (B) The cortical localization of dynein was scored in the different conditions used and is plotted in the bar graph. Error bars show the SEM (n = 25 in each of three independent experiments.