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. 2014 Oct 27;289(50):34601–34619. doi: 10.1074/jbc.M114.569244

FIGURE 6.

FIGURE 6.

NPM1 levels dictate the structure of the nucleolus. A, U2OS cells were transfected with WT NPM1 (Myc-NPM1) and mutant NPM1 (Myc-NPM1-(117–294)). Two days post-transfection, the cells were fixed and immunofluorescence-stained for the Myc tag using the 9E10 mAb and H3K9me3 by using a rabbit polyclonal antibody. Nuclei were stained with DAPI. The DAPI and H3K9me3 staining of representative cell nuclei indicated that the perinucleolar heterochromatin had become more dense and accentuated in wild type Myc-NPM1-expressing cells than in cells transfected with mutant Myc-NPM1-(117–294). B and C, exogenous expression of WT Myc-NPM1 in p53−/−,Npm1−/− MEFs restored nucleolar morphology to WT. p53−/−,Npm1−/− MEFs were transfected with Myc-NPM1, and after 2 days the cells were fixed and stained for NPM1 and fibrillarin (B) or NPM1 and H3K9me3 (C). D, failure of mutant NPM1 to restore nucleolar morphology in p53−/−,Npm1−/− MEFs to WT. Cells were transfected with Myc-NPM1-(117–294) mutant and stained for the Myc tag and H3K9me3.