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. 2011 May 6;6(5):e19596. doi: 10.1371/journal.pone.0019596

Figure 4. Cell cycle dependent interactions between Tim and mitotic entry kinases.

Figure 4

A–B) siControl and siTim transfected HCT116 cells were synchronized by double thymidine and then assayed by PI staining and FACS analysis for cell cycle profile. C) Western blot of total cell extracts of synchronize HCT116 cells after siControl or siTim at cell stages shown in panels A and B for 3, 6, 9, and 12 hrs post-release from thymidine block. Western blot with antibody to Tim, Plk1, Aurora A1, Aurora B1, Cyclin B1, CDK1, Cdh1, Cdc25, histone H3 phospho S10, hitone H3, or Actin, are indicated. D) Immunoprecipitation with anti-Tim or control IgG antibody with extracts from HCT116 cells at 0, 2, 4, 6, or 8 hrs post-arrest from thymidine block. IPs were assayed by Western immunoblot (IB) with Tim, Plk1 Aurora A, or Aurora B antibody as indicated. E) HCT116 cells were synchronized as in panel D, and extracts were subject to IP with antibody to Plk1 (middle panel) or Aurora A (lower panel), followed by Western blot with anti-Tim. Input is shown in top panel, as indicated.