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. 2013 Apr 15;24(8):1208–1221. doi: 10.1091/mbc.E12-06-0450

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© 2013 Quaresma 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 4: — Inhibition of PI3 kinase or AKT increases cytoplasmic levels and reduces nuclear levels of poly(A) RNA. HeLa cells were treated for 4 h with 20 μM LY294002, 5 μM AKT inhibitor VIII, or 100 nM rapamycin before FISH. Cells were fixed for 10 min on ice and then hybridized with biotinylated 54-mer oligo(dT). The hybridized probe was detected with Alexa 488–conjugated streptavidin. Cells were counterstained with 4′,6-diamidino-2-phenylindole to identify nuclei. (A) Representative field of the poly(A) distribution in untreated HeLa cells. Scale bar, 20 μm. (B) Bar graph of poly(A) in the nucleus or cytoplasm. Values are plotted as the percentage of the total cellular poly(A). Statistical analysis of the significance of differences between untreated and treated samples is by t test. Means are plotted with SDs. ***p ≤ 0.001.