Figure 2.

Centrosome Depletion Following PLK4 or STIL Blocking Leads to Prolonged Mitosis and Mitotic Defects

(A) Phase-contrast images of 2-day vehicle- and centrinone-treated hESCs and hiPSCs or 2 days after STIL shRNA transfection. Arrows indicate mitotic cells. Scale bars, 50 μm.

(B–E) Cell-cycle distribution of 3-day vehicle- and centrinone-treated hESCs (B) or hiPSCs (C) analyzed by FACS. Measurement of relative length of mitosis (D) or interphase (E) by live imaging of H2A-GFP hESCs after indicated time of treatment. Data are normalized to the vehicle treatment condition on day 1 (n = 2, N > 40).

(F) Immunofluorescence analyses of centromere number in 4-day vehicle- or centrinone-treated hESCs and hiPSCs. Centromeres were visualized by CREST staining (yellow), nuclei were counterstained by Hoechst (green). Scale bars, 10 μm. Panels on the right show centromere quantification and corresponding intervals of chromosome numbers (n = 2, N > 90).

(G) Quantification of viability measurement by annexin V/PI staining in 2-day vehicle- and centrinone-treated hESCs and hiPSCs.

Data are presented as mean ± SEM (^∗p < 0.05, ^∗∗p < 0.005). See also Figure S2.