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. 2012 Feb 15;11(4):807–817. doi: 10.4161/cc.11.4.19323

Figure 1.

Figure 1

MK8745 is a selective inhibitor of Aurora A kinase. (A) Chemical structure of MK8745. (B) In vitro kinase assay. (i) Using plk peptide or Histone H3 substrate and purified Aurora A Kinase with (MK), without (ND) 500 nM MK8745 or with 500 nM AZD1152 (ABI). Phospho plk1 (S137), phospho H3 (S10) and Aurora A were detected by western blot analysis. (ii) Using purified Aurora A kinase as substrate with or without MK followed by western blot analysis for phospho (T288) Aurora A. (iii) Using Histone H3 as substrate and Aurora B kinase with or without MK followed by western blot analysis for phospho histone (Ser10) H3. (iv) Western blot analysis for phospho H3 (S10), Aurora A and tubulin in lysates of HCT 116 cells treated with MK for 24 h. (v) Flow cytometry analysis of HCT cells treated with (MK) or without (ND) MK for 24 h probed for MPM2, a mitotic marker. (C) Western blot analysis of HCT p53−/− cells overexpressing p53 treated with (MK) or without (ND) probed for phospho (Ser315) p53 and p53. (D) Coimmunoprecipitation of p53 with Aurora A and not with Aurora B in HCT 116 cells. All the results shown are representative of 3–4 independent experiments.