Phospho-mimetic Mps1^8D impairs the kinetochore recruitment of BubR1 and Mad2.

<p>(A) Representative immunofluorescence images of prometaphase cells co-transfected with Mps1 shRNA and GFP vector at a 3∶1 ratio. Cells were fixed and co-stained for BubR1 (red) and DNA (blue). Scale bar represents 10 µm. (B) and (C) Representative immunofluorescence images of prometaphase cells expressing different LAP-tagged Mps1 constructs. At 36 hours after co-transfection with the Mps1 shRNA and indicated plasmids, cells were fixed and co-stained for BubR1 (red) (B) or Mad2 (C), and DNA (blue). Scale bar represents 10 µm. (D) Bar graph showing quantification of kinetochore signal of BubR1 and Mad2 in cells expressing different Mps1 constructs as indicated. Bars indicate mean ±SE from 3 independent experiments. In each experiment, 5 cells were measured (>60 kinetochores per cell). * <i>P</i><0.001 versus shMock or Mps1^WT and Mps1^8A rescue groups.