Figure 3.

Hec1 and Hice1 are components of the centrosome. (A) Double immunostaining of Hec1 and β-tubulin in U2OS. Right, line scanning shows the relative fluorescence intensity along the line connecting the spindle poles. (B) Immunostaining for Hec1 in U2OS cells stably expressing Hice1-GFP. Arrows indicate centrosome/spindle poles (A and B). (C) Isolated KE37 centrosomes were prepared by discontinuous sucrose gradient centrifugation and serial fractions were separated by SDS-PAGE and subjected to Western blotting analysis with anti-γ-tubulin, Hec1, and Hice1 antibodies. (D) Double immunostaining of purified KE37 centrosomes with anti-γ-tubulin and Hec1 or Hice1 antibody. Bar, 10 μm. DAPI staining was used to reveal chromosome/nucleus in A and B.