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. 2016 Mar 4;44(11):5105–5122. doi: 10.1093/nar/gkw113

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 6. — pancNusap1 overexpression (OE) caused cells subjected to the irreversibly differentiation condition to have the morphological and epigenetic characters of reversibly differentiated cells. (A) Genomic structure of the rat Nusap1 promoter. Bold-lined boxes show the region homologous to mouse Oip5. Gray boxes indicate predicted locations composing the Oip5 open reading frame (ORF). Lower thick arrows indicate the region and transcribed strands used for OE-experiments of ncRNA. Upper and lower vertical lines show the positions of methionines (Met) or stop codons (Stop), respectively, in each translational frame. (B) The effect of Dox-induced sense or antisense pancNusap1 OE on the Nusap1 expression level in NcAdiff cells. Gapdh was used as a control. Empty vector-introduced PC12 cells were used as a negative control. (C) Histone modification status of the Nusap1 promoter in pancNusap1-overexpressing PC12 cells under irreversible differentiation condition. (D) The effect of Dox-induced sense or antisense pancNusap1 OE on cell proliferation in NcAdiff cells. Upper diagram shows experimental scheme. Number of living cells was counted. After 7 days of differentiation of PC12 cells, NGF/cAMP were removed to allow proliferation. (E) Proportion of Ki67+ cells in antisense pancNusap1-overexpressing PC12 cells after NGF/cAMP deprivation. In (B–E), values are mean ± SD (n = 3). Statistical analysis was performed using Student's two-tailed t-tests or the Tukey-Kramer multiple comparison test. N.S. = not significant. *P < 0.05; **P < 0.01; ***P < 0.001.