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Figure 3


Fig. 3. Synchronization at S-phase causes accumulation of BAF in the nucleus. (A) Indirect immunofluorescence staining of endogenous BAF (upper panels) or exogenously expressed HA-BAF (lower panels) in non-synchronized (left panels) and S-phase synchronized (right panels) HeLa cells. Bar, 10 µm. (B) Endogenous BAF was stained in non-synchronized (control) and S-phase synchronized HeLa cells 0, 2, 4, and 6 hours after release from the S-phase block. Percentages of the cells with nucleus-positive (black bar), uniform (grey bar) and cytoplasm-positive (white bar) BAF localization patterns are shown. Number of cells examined: 297 for control, 629 for 0 hours, 548 for 2 hours, 714 for 4 hours and 599 for 6 hours. (C) Western blotting analysis using anti-BAF antibody. Whole cell extract (total), the cytoplasmic fraction (cytosol), and the nuclear fraction (nucleus) from non-synchronized (control) and S-phase synchronized (S-phase) HeLa cells were analyzed. Lactate dehydrogenase isoenzyme I (LDHI) and Lamin B were used as markers for cytosol and nucleus, respectively.





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