Figure 1
Cdc5L deficiency causes mitotic arrest and chromosome misalignment. (a and b) HeLa cells were transfected with control siRNA or two independent siRNAs against Cdc5L. (a) FACS analysis of cell cycle distribution by PI staining at indicated times post transfection. (b)The proportion of mitotic cells in total cells were analyzed by FACS using phosphorylated histone H3 (Ser-10) (p-H3)-PI staining at indicated times post transfection. (c) Selected frames from time-lapse movies of representative HeLa/GFP-H2B cells transfected with control or Cdc5L siRNA. The time on the images is in minutes. A, anaphase; M, metaphase; NEB, nuclear envelope breakdown. Scale bar, 10 μm. (d) The percentage of mitotic incompetent cells (mitotic duration >90 min) in control and Cdc5L-knockdown cells entering mitosis. Data are representative of three independent experiments, error bars indicate S.D. (e) The percentage of mitotic cells with misaligned chromosome in control and Cdc5L-knockdown cells entering mitosis. Data are representative of four independent experiments, error bars indicate S.D. (f) Complementation of red fluorescent protein (RFP)-Cdc5L in knockdown HeLa/GFP-H2B cells rescues chromosome misalignment. The Cdc5L-knockdown cells were transfected with Cdc5L siRNA-resistant expression construct or control vector. The data show the percentage of mitotic cells with misaligned chromosome in RFP-positive cells entering mitosis. Data are representative of four independent experiments, error bars indicate S.D.
