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. 2007 Sep 10;178(6):913–924. doi: 10.1083/jcb.200702058

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Copyright © 2007, The Rockefeller University Press

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Figure 3. — Cellular localization and subnuclear fractionation of GFP-UTF1 in EC and ES cells. (A) Western blot analysis of wild type (wt) EC cells and a clone stably expressing GFP-UTF1 (#1) using an antibody directed against UTF1. (B) Subnuclear fractionation of EC and ES cells, both expressing GFP-UTF1. Immunoblot analysis was performed with an antibody directed against UTF1 and the HA tag of the fusion protein. F, free-diffusing protein fraction; D, DNaseI fraction; AS, ammonium sulfate fraction; HS, high salt fraction; M, nuclear matrix fraction. (C) Confocal and transmission image of living EC cells expressing GFP-UTF1. Nucleoli are indicated by arrows. (D) Time-lapse imaging of a GFP-UTF1–expressing EC cell going through mitosis. (E) Confocal images of a mitotic GFP-UTF1–expressing cell treated with Hoechst. (F) Confocal and transmission images of GFP-UTF1–expressing ES cells grown on an STO cell feeder layer. The arrow indicates a nucleolus, and the arrowhead points to mitoticchromosomes. (G) Transmission image of AP staining of GFP-UTF1 ES cells. The underlying STO feeder cells are negative for AP activity. Bars, 15 μm.