EB1 localizes Aurora B at the centromere in a microtubule-dependent manner. (A) Loss of centromeric Aurora B is rescued by expressing siRNA-resistant EB1 (EB1siRes) but not the EB1K89E mutant (EB1K89EsiRes). Bar, 2.2 µm. (B) Loss of KNL1(Ser60) phosphorylation was rescued by expressing siRNA-resistant EB1 but not the EB1K89E mutant. Bar, 2.3 µm. (C) Quantification of immunostaining Aurora B intensities in A. *, P = 3.78 × 10−69; **, P = 2.54 × 10−75; ***, P = 1.33 × 10−35. (D) Quantification of immunostaining phospho-KNL1(S60) intensities in B. *, P = 5.23 × 10−42; **, P = 1.34 × 10−42; ***, P = 1.02 × 10−39. (E) Western blot of HeLa lysates showing endogenous and GFP-tagged EB1 and KNL1 levels (in kilodaltons). (Additional rescue experiments are shown in Fig. S2, A–C.) The height of the boxes represents the IQR. The central horizontal lines depict the median. The top whiskers represent the 75th percentile + 1.5× IQR, and the bottom whiskers represent the 25th percentile − 1.5× IQR. a.u., arbitrary unit.
