Effect of AM-D on FhkA and CBP4a localization. (A) AM-D treatment (4 h) resulted in the budding of nucleolar-localized FhkA from the nucleus. After 8 hours FhkA was detected as cytoplasmic circles adjacent to the nucleus, always on the opposite side as the centrosome. After recovery (4 hr) from AM-D treatment FhkA was still detected in cytoplasmic circles (Cir) which were not adjacent to the nucleus. (B) Enlargement of nucleus from (A). (C) In AM-D-treated cells (4 hr) the intranuclear patches detected by anti-CBP4a were diminished in size compared to those detected in untreated cells. These patches ultimately disappeared after 8 hr treatment. (D) Enlargement of nucleus from (C). Four hour treatment also resulted in detection of CBP4a in the centrosomal region (Ct) while 8 hour treatment resulted in the presence of cytoplasmic particles (CP). Treatment for 4 hours followed by a 4 hour recovery period resulted in only a partial restoration of nucleoplasmic CBP4a. Scale bar represents 2 μm. CBP4a budding (B), cytoplasmic particles (CP), centrosomes (Ct), and nucleoli (No) are indicated. At least four independent replicates were performed, all of which showed the same result.