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Fig. 5. Inhibition of Aurora-A expression by RNA interference shut down serine 353 phosphorylation. HeLa C1 cells expressing GFP-centrin (Piel et al., 2001) were transfected with Aurora-A RNAi or control scrambled RNAi. Immunofluorescence staining was performed with Aurora-A monoclonal antibodies (A), with SE96 anti-serine 353-phosphorylated CDC25B polyclonal antibody (B), or with a CDC25B polyclonal antibody. Cells were also stained with DAPI (blue). (D) Lysates from HeLa cells treated as above were subjected to western blot analysis with anti-Aurora-A and anti ß-tubulin as a loading control. (E) Quantification of immunofluorescence signal was performed using Metamorph software on images taken with a CoolsnapHQ camera.





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