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Figure S5. hSgo2 is not essential for centromere cohesion and localization of chromosomal passenger proteins. (A) Chromosome spreads depicting normal and defective cohesion defects (parallel and separated). These criteria were used to examine chromosome morphologies in cells transfected with different siRNAs. 36 h after transfection with control, hSgo1, hSgo2, and MCAK siRNAs, cells were treated with nocodazole and harvested by shake-off 3 h later. Chromosome spreads were prepared and parallel samples were processed for staining to confirm >90% depletion of target proteins. (bottom) The distributions of normal, separated chromatids or centromeric separation were scored. (B) Ratios of Mad1 staining intensities at kinetochores of unaligned versus aligned chromosomes in control and hSgo2 siRNA-transfected cells (n > 50; P = 0.86). (C) HeLa cells transfected with control and hSgo2 siRNAs were costained with hSgo2 and Aurora B (top), INCENP (middle), and survivin (bottom). Chromosomes were stained with DAPI. Images are from maximum projections. Exposure times between control and hSgo2 siRNA samples were identical.