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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 3: — Phosphorylation of UHRF1 requires ccna2. (A) Experimental design. Embryos obtained from a cross of heterozygous ccna2^hi2696 mutants were injected (arrow) with a construct containing UHRF1-EGFP driven by the heat shock (hsp70l) promoter. Mutants (red) were separated from wild-type (tan) siblings based on phenotype at 3 dpf, heat-shocked at 4 dpf, and stained for the presence of phosphorylated UHRF1. (B) Immunofluorescence of 4-dpf wild-type (WT) and ccna2^hi2696 embryos shows that ccna2^hi2696 mutants do not have phosphorylation of UHRF1 despite overexpression of UHRF1, as seen by the GFP, due to mosaic expression from the Tg(hsp70I: UHRF1-EGFP) transgene. Mosaic expression is obtained by this method, so not all nuclei are labeled with GFP. Scale bar: 25 μm.