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. 2017 Dec 19;7:17761. doi: 10.1038/s41598-017-18087-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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NS1 of Influenza A/Shantou/602/06 (H3N2) induces nucleolar stress depending on its association with NCL. (A) Immunofluorescence staining of NS1 Mutant (218–230 aa truncated mutant) (Green), or NCL (Red) in A549 cells. (B) Lysates from mock-transfected, NS1-transfected or NS1 Mutant-transfected A549 cells were subjected to immunoprecipitation using anti-Flag antibody and immunoblotted with anti-NCL or anti-Flag antibodies. 5% cell lysate was served as input. (C) Cell lysates harvested 36 h post-transfection were processed for western blot with p53 antibody. β-actin was used as the loading control. (D) RT-qPCR analysis of pre-45S rRNA and p21 (E) in transfected A549 cells. Results show the mean of the three experiments ± SD and are normalized to the β-actin gene. *p < 0.05, **p < 0.01, ***p < 0.001 (t-test).