Figure 4.
PP2A-AαP179R/+ Cells Exhibit More Robust Centrosome Clustering
(A) Schematic of centrosome amplification induction by doxycycline (dox)-inducible Plk4 overexpression.
(B and C) Plk4-inducible cells were treated with dox and fixed and analyzed by immunofluorescence. (B) Representative bipolar (top) and multipolar (bottom) anaphases. (C) Multipolar anaphase incidence in cells with >4 centrin-1 foci is plotted.
(D–F) Plk4-inducible cells were treated with dox and imaged live (D) GFP-H2B (top) and differential interference contrast (bottom) montage. Time (minutes) relative to nuclear envelope breakdown is indicated. (E) Incidence of multipolar cell division is plotted. (F) A box-and-whisker plot of mitotic duration in dox-treated cells undergoing bipolar cell divisions. Whiskers indicate the 5th–95th percentile range, and circles indicate cells outside of this range. Result is representative of three experiments.
(G and H) Plk4-inducible cells were treated with dox and imaged on O- and Y-shaped fibronectin micro-patterns. (G) Representative images of interphase cells. (H) Incidence of multipolar anaphases. n, total cells analyzed.
(C and E) Mean ± SEM from three experiments. Scale bars, 5 μm. ***p < 0.0005, **p < 0.005; ns, not significant (p > 0.05) Student's t test.
See also Figure S2.