FIGURE 5:

Nek2 activity is required for phospho-S33/S37/T41 reactivity at spindle poles (A, C) U2OS cells were synchronized in mitosis by double-thymidine block and release, transfected with HA-KD Nek2 or HA-WT Nek2, and immunostained with antibodies as indicated. For presentation of control spindles and different treatments, images were taken at identical exposure times and identically contrast enhanced for each stain. Scale bar, 10 μm. (B, D) Levels of β-catenin (B) and phospho-S33/S37/T41 immunofluorescence (D; AU, arbitrary units) at spindle poles of untreated U2OS cells or cells treated as described in A and C. Error bars, SEM of ≥15 spindle poles for B and SEM of ≥15 spindle poles for D per experiment from two experiments; ***p < 0.001. Original unmodified images taken at identical exposure times were measured for controls and transfected cells.