UNC-45a depletion results in delayed cytokinesis and multinucleated cells. (A) Visualization of actin and nuclei (DAPI) in UNC-45a knockout (KO) cells. Bars, 20 µm. (B) Quantification of multinuclear WT and UNC-45a knockout cells. Graph displays percentage of multinuclear cells; means ± SEM, WT, n = 768 and knockout, n = 269 cells. (C) Phase-contrast, time-lapse images on cleavage furrow ingression rates of WT and UNC-45a knockout cells. Time 0 refers to the onset of anaphase. Location of the kymograph is indicated by the orange, dashed line. Orange arrows mark the direction of ingression in the kymograph. Bars, 20 µm. (D) Furrow ingression rate quantification from the phase-contrast, time-lapse data. WT, n = 31; UNC-45a knockout, 20 cells from three experiments (Student’s t test). (E) SIM images on cleavage furrow of WT and UNC-45a knockout cells with phalloidin-labeled actin filaments (magenta) and NM-IIA rods (green). Magnified views of the ingressing furrow (orange boxes) are displayed on the right. Bars, 5 µm. (F) Quantification of NM-IIA stack lengths in the cleavage furrow of WT and UNC-45a knockout cells. WT, n = 6 (1,928 filaments); knockout, n = 5 (4,838 filaments) cells. Data are represented as means ± SEM (Mann–Whitney test).