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. 2018 Jun 6;8:8680. doi: 10.1038/s41598-018-27008-w

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© The Author(s) 2018

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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RPS3 interacts with α-tubulin and modulates the organization of α-tubulin. (A) Organization of α-tubulin in control and shRPS3-4-transduced HeLa cells. HeLa cells were transduced with the control virus or shRPS3-4 and immunostained with antibodies against RPS3 and α-tubulin. Control, n = 114; shRPS3, n = 134. Experiments were repeated three times. (B) α-Tubulin assembly in GFP- or RPS3-GFP-transfected cells. HeLa cells were transfected with an expression construct for GFP or RPS3-GFP and immunostained with an anti-α-tubulin antibody. Control, n = 24; RPS3 = 49. Experiments were repeated two times. (C) Expression of RPS3-GFP in RPS3-depleted cells rescues abnormal α-tubulin assembly. HeLa cells were transduced with shRPS3 and transfected with an expression construct for GFP or RPS3-GFP the next day. Cells were further incubated for 48 h and immunostained with an anti-α-tubulin antibody. Control, n = 20; RPS3, n = 51. Experiments were repeated three times. (D) RPS3 interacts with α-tubulin in G2/M phase. Cell extracts were prepared from G2/M-arrested HeLa cells, and immunoprecipitation was performed with an anti-α-tubulin antibody. Precipitants were subjected to western blotting with an anti-RPS3 or anti-α-tubulin antibody. *Non-specific band.