Figure 6.

Proteins identified by the MS- and the HPA-screen display asymmetric centrosome localization. (A) C14orf145, (B) Albatross, and (C) PRICKLE3 stain a single centrosome/centriole at the G1/S-phase of the cell cycle in U-2 OS cells as compared with the centrosomal marker protein γ-tubulin (Cy3). To distinguish between mother and daughter centriole association, the axoneme-extended mother centriole were stained with anti-acetylated tubulin (Cy3) in ciliated hTERT-RPE1 cells. This indicates that C14orf145 and Albatross localize to the mother centriole. (D) Antibody staining of GFP–C3orf34 in HeLa cells at the G1/S-phase and MPHOSPH9 in RPE cells at the G0 phase. Co-staining with the mother centriolar marker protein ODF2 (Cy5) indicates that GFP–C3orf34 associate with the mother centriole. Co-staining with anti-acetylated tubulin (red) in ciliated hTERT-RPE1 cells suggest that MPHOSPH9 (green) localize proximal at the mother centriole and distal and proximal at the daughter centriole. DNA was stained with DAPI, yellow indicates coincidence of green and red signals. Bars, 5 μm. (E, F) Interpretation of the staining patterns of tubulin (red) and asymmetrically localized candidate proteins (green) in duplicated centrosomes and basal bodies.