FIGURE 8.

D-box mutant preventing Mps1 degradation causes centrosome amplification and mitotic abnormalities. 293T cells were depleted from the endogenous Mps1 by using pSUPER.retro siRNA as described before and then transfected with GFP-tagged Mps1 wild type or D-box mutant constructs. 72–96 h post-transfection, cells were subjected to immunofluorescence by using anti-γ or anti-α antibody. A, representative images and bar graph shows that quantitation of frequency of cells with more than two centrosomes. ∼200 interphase cells were assessed at each treatment. Three independent experiments were performed. B, representative images and bar graph show the frequency of cells with abnormal spindles. ∼200 mitotic cells were assessed at each treatment. Three independent experiments were performed. C, curve graph shows the cell cycle distribution. Three independent experiments were performed. P, prophase; P-M/M, prometaphase/metaphase.