(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.