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Figure 6


Fig. 6. The persistence of cohesin on chromosome arms in Aurora B depleted cells does not depend on Sgo1. (A) In order to visualise cohesin in mitosis, expression of Scc1-9xMyc was induced by applying Doxycyclin (1 µg/ml for 48 hours). Cells were spread onto glass slides using hypotonic conditions, fixed and stained with antibodies against Myc, Sgo1 and the autoimmune serum CREST to detect centromeres. Sgo1 and Aurora B were depleted by siRNA transfection. 100 µm Hesperadin was used to inhibit Aurora B activity. Bar, 10 µm. (B) Number of cells (%) showing only centromeric cohesin versus cells which display cohesin also on chromosome arms. (C) Number of cells (%) showing paired versus single sister centromeres. (D) Images from a movie of HeLa cells stably expressing Sgo1-GFP. Centromeric enrichment of Sgo1-GFP is lost when Hesperadin (100 nM) is added to the medium (t=0). Chromosomes were visualised with Hoechst 33342 (0.2 µg/ml). Movies were taken on an Olympus DeltaVision microscope (63x objective, five slices every 2 minutes, 2x2 binning). Projected images at times indicated (in minutes) are shown.





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