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Figure 1


Fig. 1. Characterisation of model system. HEK293 cell lines stably transfected with Aurora transgenes were induced with tetracycline then analysed by immunoblot, immunofluorescence and immunoprecipitation kinase assays. (A) Immunoblot showing that the anti-Aurora A, B and C antibodies are monospecific for Aurora (Ar) A, B and C respectively. DN, Aurora kinase D-N mutant; KR, Aurora kinase K-R mutant; WT, wild type. (B-D) Immunoblots probed with antibodies against Aurora proteins, the Myc-epitope tag and phosphorylated Histone H3 (S10), showing tetracycline induced expression of Aurora transgenes and effects on H3 phosphorylation. (E) Immunofluorescence images showing localisation of exogenous Aurora proteins. Bar, 5 µm. (F,G) Immunoprecipitation kinase assays showing that the Aurora kinase mutants are catalytically inactive.





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