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. 2004 Nov;78(21):11487–11505. doi: 10.1128/JVI.78.21.11487-11505.2004

FIG. 4.

FIG. 4.

1A-DBD, 2A-DBD, and 3A-DBD bind metaphase chromosomes. Metaphase chromosomes were isolated from 293, 293/DBD, 293/EBNA1, 293/1A-DBD, 293/2A-DBD, and 293/3A-DBD cells stalled in mitosis by Colcemid treatment. Indirect immunofluorescence was performed by using the K67.3 rabbit polyclonal antibody against the DBD of EBNA1. Images of individual layers (40 z-sections of 100 nm each) were captured using a ×100 objective, with ×1.5 optical enhancement, and deconvolved by using softWoRx. The cell line used for metaphase chromosome isolation is indicated above each panel. All three proteins were observed to localize to metaphase chromosomes in discrete punctate spots that resemble those observed previously with wild-type EBNA1 or HMGA1a-DBD (48).