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Fig. 7. Silencing of BBS6 by RNAi in COS-7 and NIH 3T3 cells produces multi-nucleated and multi-centrosomal cells, and cytokinesis defects. (A) DIC images of COS-7 and NIH 3T3 cells 9 days after transfection with BBS6 siRNA expression vector (pSilencer-BBS6 or pSuper-BBS6, respectively) show interconnected cells with long intercellular bridges as a result of defective cytokinesis. Immunocytochemistry with a ${gamma}$ -tubulin antibody (red) and DAPI staining of nuclei (blue) show that BBS6 silencing causes bi-nucleated and multi-centrosomal phenotypes in both COS-7 and NIH 3T3 cells. The merged images (right panels) show two nuclei in single cells. (B) Microtubule organization is not significantly altered during cell cycle progression in COS-7 cells transfected with pSilencer-BBS6 (upper panels) compared with cells transfected with pSilencer empty vector (lower panels). Cells were stained with antibodies to ${alpha}$ -tubulin (red), ${gamma}$ -tubulin (green) and counterstained with DAPI (blue). Only merged images are shown; yellow indicates overlapping ${alpha}$ - and ${gamma}$ -tubulin signals.

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