All stages of mitosis are impeded by CA-WASp expression. (A) Time-lapse images of HT1080 cells expressing fluorescent Histone 2B showing nuclear envelope breakdown (NEB), anaphase onset (Ana), and the start (Fur) and completion (End) of furrowing. Bar = 10 μm. (B) Timing of NEB to anaphase, (C) anaphase to furrow initiation, and (D) furrow duration; n > 200 cells over at least 4 independent experiments. (E) Chromatid speed during anaphase (mean ± SEM, n = 14 control and n = 15 CA-WASp). (F) Peak chromatid speeds. (G) Furrow closure speed (mean ± SEM, n = 16 for control and CA-WASp). (H) Peak furrow closure speeds. Imaging used a Zeiss Axiovert 135 microscope fitted with an environmental chamber at 37°C with 5% CO₂. Cells were cultured in phenol red–free DMEM with 10% FCS. Fluorochromes were mCherry-H2B and eGFP-WASp. A Hamamatsu ORCA-ER CCD camera was used, and acquisition was controlled with Volocity software. Image analysis used ImageJ.