Figure 6.

CPC phase separation and MT bundling are important for formation of bundled structures on the anaphase spindle.A, experimental design to visualize spindles in the first mitosis after knockdown and replacement of Borealin by siRNA in HeLa cells. B, fluorescence images of HeLa cells with endogenous Borealin replaced with WT or indicated Borealin mutants and stained for α-tubulin and DAPI. Maximum intensity projections of representative images are shown; scale bar represents 5 μm. Experiment was repeated three times, and the different colors in the quantifications represent experimental replicates; data were normalized between experimental replicates by dividing the mean values of intensity of signal from MTs by the maximum of the total intensity within each experimental replicate; CEN-Borealin^wt (n = 28, 38, 32); CEN-Borealin^LLPS (n = 26, 28, 42); and CEN-Borealin^MTBM (n = 26, 30, 38). p^wt-LLPS = (0.015, 0.00022, 0.0058); p^MTBM = (1.77× 10⁻⁶, 6.9 × 10⁻⁵, 3.3 × 10⁻⁸). For statistical analysis, Welch's t test with Bonferroni correction was applied; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. CPC, chromosomal passenger complex; DAPI, 4′,6-diamidino-2-phenylindole; MT, microtubule.