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. Author manuscript; available in PMC: 2015 Aug 21.
Published in final edited form as: Mol Cell. 2014 Jul 17;55(4):578–591. doi: 10.1016/j.molcel.2014.06.016

Figure 6. The Cep192-organized AurA-Plk1 cascade is essential for centrosome maturation and bipolar spindle assembly in mammalian cells.

Figure 6

(A) W-blots of lysates and AurA immunoprecipitates from HeLa cells transfected with control siRNA (Mock) or Cep192 siRNA #1. In lanes 1 and 2, cells were synchronized in mitosis by a thymidine-nocodazole block. IgG h.c, heavy chain of IgG.

(B) Representative IF images of cells treated with control siRNA (Mock) or Cep192 siRNA #2. Cells were transfected with an empty vector or with siRNA-resistant cDNAs encoding the indicated HA-tagged hCep192 proteins. Insets show higher-magnification views of the centrosome regions. Scale bars, 10 µm.

(C) IF of Plk1 and centrin in cells transfected with control siRNA (Mock) or Cep192 siRNA #1.

(D) MT regrowth assay in Cep192 siRNA #2-treated cells expressing the indicated HA-tagged hCep1 92 proteins.

(E) Quantification of the centrosome volume in [(B), HA signal] and of the IF intensities of the centrosomal γ-tubulin, kinesin-5 in (B) and α/β-tubulin in (D) (mean +/− SD). At least 50 MT structures were analyzed for each parameter in each of the three independent experiments. *P<0.001.

(F) Quantification of the spindle assembly defects in cells in [(B), panel 3] (mean of three independent experiments +/− SD).

See also Figure S7.