Fig. 7.

Subcellular localization of pS894-INCENP during mitosis. (A, B, and D) Synchronized HeLa-S3 cells at the indicated stages of the cell cycle were immunostained with anti-pS894 (red), and counterstained with anti–α-tubulin (green in A), anti-INCENP (green in B), and anti-Aurora-B (green in D). DNA was stained with Hoechst 33258 (blue). Insets in D show magnified images of the kinetochore (in prometaphase and metaphase), central spindle (in anaphase and telophase), and midbody (in late telophase and cytokinesis). Scale bar, 10 μm. (C) Peptide competition assay. After the preincubation of anti-pS894 antibody with phosphorylated or non-phosphorylated S894 peptides, HeLa-S3 cells synchronized at metaphase were immunostained with the preincubated antibody (red) and counterstained with anti-INCENP (green) antibody. Scale bar, 10 μm. (E) Nuclear and cytoplasmic localization of pS894-INCENP during G2 phase. Arrow and arrowhead show a G2 phase cell and a prophase cell, respectively. Scale bar, 10 μm.