Fig. 1.

GAK is required for efficient chromosome alignment. a Hela cells were transfected with six different siRNAs targeting GAK or with an siRNA targeting GAPDH as a control. Twenty-four hours after transfection cells were re-transfected and 48 h after the first transfection cells were fixed, DNA was stained with DAPI, and the percentage of mitotic cells with misaligned chromosomes was scored in each condition. b Hela cells were transfected as in (a) with indicated siRNAs, and 48 h after the first transfection cells were lysed and GAK protein levels were determined by western blot. c, d Hela (c) or U2OS cells (d) were treated as in (a), and the average number of misaligned chromosomes was determined (c) or the percentage of mitotic cells with misaligned chromosomes was scored (d). e Hela cells were treated as in (a), but 48 h after transfection cells were fixed and stained for CLIP-170 and p150glued to visualize unattached kinetochores and DAPI was used to stain the DNA. All graphs are averages of three independent experiments with 50 cells scored per condition per experiment. Error bars represent standard deviations. Scale bar in (e) indicates 5 µm