Figure 4.

Dynein is necessary for the localization of BRCA2 to the centrosome. (A) Western blot analysis of lysates prepared from HeLa S3 cells transfected with siRNA targeting dynein 1 heavy chain 1 (DHC1) (DHC1-1 and DHC1-2 siRNA) or control siRNA or from non-transfected cells (siRNA-free) using anti-DHC1 and anti-β-actin antibodies. (B) Representative immunofluorescence microscopy of HeLa S3 cells treated as in (A) using anti-γ-tubulin (red) and anti-BRCA2 (green) antibodies. Nuclei were stained with Hoechst 33258. (C) Quantification of BRCA2 localization to the centrosome in DHC1 siRNA-transfected and transfection control cells. (D) Staining of control (DMSO-treated) and EHNA-treated HeLa S3 cells with antibodies against anti-centrin 3 (green) and anti-BRCA2 (red) antibodies. Nuclei were stained with Hoechst 33258. The boxed areas are shown at higher magnification in the right panels.