Fig. 8. hSMC1 is mislocalized along chromosome arms with CENP-A specifically during mitosis. (A) hSMC1 was immunolocalized in HeLa cells that were transiently transfected with HA-tagged CENP-A 48 hours prior to arrest in mitosis. Cells overexpressing CENP-A-(HA), detected with anti-HA antibody, demonstrated anti-hSMC1 mislocalized along the arms of chromosomes (cell on right). Control untransfected cells within the same preparation (cell on left) demonstrated the characteristic centromere-kinetochore-exclusive staining of anti-hSMC1 during mitosis. DNA was counterstained with DAPI. (B) CHO cells were grown and processed on coverslips to examine ectopic and centromeric hSMC1 at all stages of mitosis. hSMC1 was observed along the lengths of chromosomes as early as prometaphase. SH-CREST was used to detect the mistargeted CENP-A. (C) Centromeric and non-centromeric anti-hSMC1 staining was redistributed to perichromosomal foci during late anaphase. Centromeric and non-centromeric SH-CREST staining, however, was observed at all stages. Bars, 10 µm.