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. 2010 Dec 1;21(23):4212–4226. doi: 10.1091/mbc.E10-04-0287

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© 2010 by The American Society for Cell Biology

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 2. — Localization of endogenous RRP1B is predominantly nucleolar and sensitive to RNAse treatment. (A) In interphase U2OS cells, endogenous RRP1B (red) is predominantly nucleolar (arrow). The metaphase cell in this field shows perichromatin accumulations of RRP1B (arrowhead). (B) The telophase cell in this field shows that RRP1B is found initially in bright foci that likely represent prenucleolar bodies (PNBs; hashed arrows) at the end of mitosis, and later in mature nucleoli (arrows). (C) Endogenous RRP1B stained with anti-RRP1B antibodies (green) in U2OS cells colocalizes with the nucleolar RNA signal (arrow) visualized by Pyronin Y staining (red). (D) Treatment of live cells with RNAse before fixation results in a near complete loss of both the RNA and anti-RRP1B, but not the DNA signal. (E) HeLa cells show a similar colocalization in nucleoli (arrows) of RRP1B (green) and Pyronin Y-stained RNA (red) and also demonstrate the sensitivity of RRP1B to RNAse treatment (F). Scale bars are 15 μm.