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Figure 3


Fig. 3. Localization of fluorescent dots of EBNA1 on mitotic chromosomes. (A) Cells were processed for immunofluorescence analyses with anti-HA antibody. Chromosomes are counterstained with DAPI. Note that fluorescent dots of EBNA1 (red) associate with prophase (left panel) and anaphase (right panel) chromosomes. (B) Dual-color immunofluorescence analyses to visualize the localization of EBNA1 (red) and CENP-C (green) on mitotic chromosome spreads of hyptonically swollen cells. Higher magnification images demonstrating paired EBNA1 dots symmetrically localizing on sister chromatids are shown in the accompanying sub-panels. (C) Simultaneous FISH and immunofluorescence to visualize EBV genomes (FISH) and HA-tagged EBNA1 on a mitotic chromosome spread. Merged images of FISH and immunofluorescence signals are also shown. Arrows indicate symmetrical localization on sister chromatids. Bars, 5 µm. (D) Distribution of EBV genomes on mitotic chromosome spreads of Akata cells. EBV genomes were detected by FISH analyses with an EBV genome-derived FISH probe. Chromosomes were counterstained by propidium iodide. EBV genomes (yellow signals) that symmetrically localize on sister chromatids are indicated by arrowheads. Bars, 5 µm.





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