Figure 7.
ARH localizes sequentially to the nuclear envelope, kinetochores, spindle poles, and midbody during mitosis. At the onset of prophase (A) ARH (red) colocalizes (yellow) with the dynactin complex protein p150glued (green) at the nuclear envelope. During prometaphase ARH colocalizes with dynein IC (DIC; B) and with a centromere marker (C) at the kinetochores. At the onset of metaphase (D) ARH (red) is found on both kinetochores and spindle poles labeled with γ-tubulin (green). During metaphase (E) ARH colocalizes with γ-tubulin at spindle poles but at the end of anaphase/beginning of telophase (F), ARH is not detectable at the spindle poles labeled with γ-tubulin (arrowheads). During cytokinesis (G and H), ARH and dynein IC (DIC) are found at the midbody. Rat-1 cells (A–F) or HeLa cells (G and H) were fixed with methanol followed by acetone, stained with ARH, markers for the mitotic apparatus or midbody, and DAPI (blue) to detect chromosomes. Bar, 6.5 μm.