Figure 6.

Relief of NTE-Mediated Autoinhibition Is Required for Activity

(A and B) Stable HeLa T-rex lines expressing MYC-GFP-Mps1-WT and mutants were treated with siMps1, synchronized in mitosis, fixed, and immunostained as indicated. The scale bar represents 5 μm. Quantification of pMELT (A) and Mad2 (B) intensities is shown. ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001.

(C and D) Cells were treated as in (A) but fixed after overnight nocodazole treatment and stained with the indicated antibodies. The mitotic index is shown; n = 3. Cells counted ≥100 per cell lines per experiment.

(E) Time-lapse microscopy of cells expressing indicated constructs. Mitotic timing is quantified. ≥50 expressing cells were counted for each condition. n = 2. ^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001.

See also Figure S6 and Movies S1, S2, S3, and S4.