Figure 2

Aurora kinase inhibitory VX-680 suppresses cell growth and induces apoptosis in TSCC cells. Cells were maintained at DMSO (served as a control) or VX-680 for 48 h. (a and b) VX-680 inhibits Aurora kinase and leads to defects in mitotic spindles. Cells were subjected to immunofluorescence staining with α-tubulin (green), Aur-A (red, original magnification × 600) or pHistone H3-Ser10 antibodies (green, original magnification × 200). DAPI (blue) was used to visualize the nuclei. (a) Quantification showed the percentage of the abnormal spindles assessed as monopolarity of three independent experiments. In each experiment, at least 150 randomly chosen spindles were counted. (b) Histogram indicated the number of pHistone H3 positive cells counted in five randomly selected fields from three independent experiments. Error bars indicated the SD. *p < 0.05, **p < 0.01, compared to control. (c-e) VX-680 suppresses cell growth and induces apoptotic cell death. (c) Cell survival rates were measured by MTT assay, *p < 0.05, ***p < 0.001. (d) Representative immunofluorescent images of apoptotic cells were stained with Annexin V (green), PI (red) and DAPI (blue). Histogram represented the percentage of Annexin V or PI positive staining cells of three independent experiments. (e) Cell apoptosis was analyzed by Western blot with indicated antibodies. GAPDH was used as a control.