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. 2012 Jan 1;23(1):59–70. doi: 10.1091/mbc.E11-06-0487

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© 2012 Chu et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — Phosphorylated endogenous Uhrf1 is cytoplasmic. (A) α-pS661 detects both overexpressed and endogenous phosphorylated UHRF1 in zebrafish. Top, GFP signal in hepatocytes of Tg(fabp10:UHRF1-GFP); middle, immunofluorescence with α-pS661; bottom, merge with DAPI. The gut and liver are labeled. Magnification: 20×; scale bar: 250 μm. (B) Sections of 5-dpf zebrafish expressing GFP-tagged UHRF1 under the control of the hepatocyte specific promoter (Tg(fabp10:UHRF1-EGFP)). Left, GFP expression in hepatocytes; right, immunofluorescence with α-pS661 (top) or preimmune (bottom). Confocal images taken with identical parameters. Scale bar: 50 μm. (C) Left, immunofluorescence of enterocytes with α-pS661 or preimmune. Right, merge with DAPI. Inset, 63× magnification of endogenous phosphorylated UHRF1 in the zebrafish gut with predominant nuclear localization. Confocal images taken with identical parameters. Scale bar: 50 μm. (D) Endogenous UHRF1 is phosphorylated in early development as seen with immunofluorescence on cryosections at 50% epiboly. Bottom, left, immunofluorescence on cryosections of embryos at 50% epiboly with preimmune sera; middle, DAPI alone; right, merge. Top, left, immunofluorescence with α-pS661; middle, DAPI alone; right, merge. Magnification: 63×; scale bar: 25 μm.