Fig. 3. CENP-C is mistargeted with CENP-A during mitosis. To determine if CENP-C is recruited to non-centromeric regions of chromatin to which CENP-A has been mistargeted, HA-tagged human CENP-A cDNA was transiently transfected into HeLa cells, and CENP-C was immunolocalized using a specific antiserum. Cells were arrested in mitosis with Colcemid for 
3 hours following 48 hours of CENP-A-(HA) ectopic expression and harvested by mitotic shake-off. (A) Control cells, in which CENP-A was not overexpressed, demonstrated anti-CENP-C antibody staining exclusively at centromeres. A low level of anti-HA antibody background staining was observed. DNA was counterstained with DAPI. Bar, 10 µm. (B) In transfected cells, anti-CENP-C antibody was found to colocalize along the lengths of chromosome arms with ectopic CENP-A, detected using anti-HA antibody. Bar, 10 µm. (C) Higher magnification view of a chromosome from panel B. Bar, 1 µm.
