Supplemental Material

This article contains the following supporting material:

• Figures 1-3 - Figure 1. Aurora B-RNAi reduces phospho-CENP-A staining more efficiently than phospho-histone H3 staining. HeLa S3 cells treated with control siRNA oligonucleotides or siRNA oligonucleotides targeting the Aurora B transcript were fixed and stained with antibodies against Aurora B and phospho-histone H3 (left panel) or Aurora B and phospho-CENP-A (right panel). Phospho-CENP-A-staining is reduced to nearly undetectable levels whereas phospho-histone H3 staining is not. Scale bar, 10 μm.

  Figure 2. HeLa S3 cells were transfected with GFP-INCENP^1-58 and simultaneously treated with siRNA oligonucleotides targeting INCENP and Aurora B (upper panel), INCENP and Aurora C (middle panel) or INCENP, Aurora B and Aurora C (bottom panel). Cells were stained with anti-GFP and anti-Aurora B antibodies. All immunofluorescence images are representatives of three independent experiments. Bar, 10 μm.

  Figure 3. Aurora B kinase Activity Is Not Required for Maintenance of the Chromosomal Passenger Proteins at the Centromere. HeLa S3 cells were treated either with DMSO (control, top panel) or with 10 μM ZM447439 for 1 h, fixed and stained with antibodies against phospho-CENP-A (green) and Aurora B, Survivin or INCENP (red), respectively. Bar, 10 μm.