Figure 3

Gene expression analysis in both HCT-wt and HCT-p53KO cells, presence of supernumerary centrosomes following Aurora-A/STK15 ectopic overexpression. A) Real time RT-PCR relative quantification shows a two fold expression level of Aurora-A/STK15 gene in untreated HCT-p53KO cells. B) Western blot, upper panel, by anti-STK15 antibody shows increased protein levels of Aurora-A/STK15 in HCT-STK15 and p53KO-STK15 cells. Real time RT-PCR, lower histogram, shows increased expression levels of Aurora-A/STK15 following infection of a retroviral vector encoding the full-length cDNA for Aurora-A/STK15. C) Centrosome analysis by γ-tubulin detection (green), nuclei stained with DAPI (blue). Arrowheads indicate abnormal centrosomes in HCT-STK15 and HCT-p53KO-STK15. D) Graphs summarize the percentage of cells with one, two or more than two centrosomes in untransfected or STK15 overexpressing cells.