Figure 6. The CNN1 domain is essential... | Rockefeller University Press

Figure 6.

$Figure 6. The CNN1 domain is essential for targeting AKAP450 and p150glued to the centrosome. (A) Protein fractions of centrosomes purified from nocodazole-arrested tagAKAP-wt (left) or tagAKAP-cnn1lox (right) cells were immunoblotted with antibodies against centrin-1 and specific PCM components. Signal intensities in fractions 3 and 4 were normalized against the centrin signal in the same fraction. Table shows the percentage of each PCM component in tagAKAP-cnn1lox centrosomal fractions compared with tagAKAP-wt fractions. Whole cell extract (WCE) represents 0.15% of total cell extract; 50% of final centrosome pellet was loaded in each fraction. (B) AKAP450 coimmunoprecipitates with p150glued and CDK5RAP2 (CDK5R2) from mitotic extracts of nocodazole-arrested HeLa cells. A rabbit IgG mix was used as negative control. Input lane shows cytoplasmic extract before immunoprecipitation (IP). wb, Western blotting.$

The CNN1 domain is essential for targeting AKAP450 and p150^glued to the centrosome. (A) Protein fractions of centrosomes purified from nocodazole-arrested tagAKAP-wt (left) or tagAKAP-cnn1^lox (right) cells were immunoblotted with antibodies against centrin-1 and specific PCM components. Signal intensities in fractions 3 and 4 were normalized against the centrin signal in the same fraction. Table shows the percentage of each PCM component in tagAKAP-cnn1^lox centrosomal fractions compared with tagAKAP-wt fractions. Whole cell extract (WCE) represents 0.15% of total cell extract; 50% of final centrosome pellet was loaded in each fraction. (B) AKAP450 coimmunoprecipitates with p150^glued and CDK5RAP2 (CDK5R2) from mitotic extracts of nocodazole-arrested HeLa cells. A rabbit IgG mix was used as negative control. Input lane shows cytoplasmic extract before immunoprecipitation (IP). wb, Western blotting.