Figure 3
Nuclear Aurora-A is crucial for H3T3-ph in early mitosis. (a, b) RPE1 cells were treated with inhibitors for Aurora-A (MLN, 50 nM), Aurora-B (Hes, 100 nM) and Plk1 (BI2536, 100 nM) for 2 h. The H3T3-ph was examined in cells in prophase (a) and prometaphase (b). (c, d) Quantification of the intensity of H3T3-ph signal in prophase (c) and prometaphase (d) cells, n=100. (e, f) The H3T3-ph was stained in the Aurora-A wild-type (WT) and knockout (KO) mouse embryonic fibroblast cells in prophase (n=97) (e) and prometaphase (n=107) (f). (g) Quantification of the H3T3-ph intensity in e, f. (h) Endogenous Aurora-A was knocked down by doxycycline-induced shRNA (+dox), while shRNA-resistant green fluorescent protein-tagged WT, kinase deficient (KD) Aurora-A and nuclear export signal (NES)-Aurora-A were transfected in Hela cells. H3T3-ph is examined by immunostaining. (i) Quantification of the H3T3-ph intensity in h. n=60. (j) Typical image (above) and ratio (below) of anaphase lagging chromosomes in NES-Aurora-A-transfected cells. n=90. Three or four independent experiments were performed. Data are represented as mean±s.e.m. *P<0.05 (Student’s t-test); **P<0.01 (Student’s t-test); ***P<0.001 (Student's t-test), DAPI, 4,6-diamidino-2-phenylindol; DMSO, dimethyl sulfoxide; NS, not significant. Scale bar: 10 μm.
