Loss of hKID and KIF4A induces irregular chromosome oscillations. HeLa CenpA-EGFP cells were transfected with 50 nM siRNA targeting hKID, KIF4A, or both and imaged 40 h pt. Three independent live-cell imaging experiments were performed for each siRNA treatment. Kinetochore behavior and dynamics were determined using MaKi software. (A) Interkinetochore distance was analyzed from cells of three independent experiments. Columns represent mean + SD. (B) Time-lapse movie frames of kinetochore pairs of control (mock transfected), hKID, KIF4A, and KK siRNA-transfected cells. Boxes show enlarged views of kinetochore pairs. Bar, 2 µm. (C) Oscillatory movements of selected sister pairs after different siRNA treatments were plotted against time (in frames; one frame is 7.5 s). Shown are the x-coordinates of each sister relative to the metaphase plate. Arrows indicate irregular breathings. ***, Student’s t test value P < 0.001.