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. 2019 Mar 4;9:3352. doi: 10.1038/s41598-019-39733-x

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© The Author(s) 2019

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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Knockdown of MYB family proteins and p95HER2 upregulation increase TIMP2 expression in MCF-7 cells. (A–C) Relative protein level of MYB (A), MYBL1 (B) and MYBL2 (C) after knockdown of MYB and MYBL1 (vector cells) or MYBL2 (p95HER2 cells). 48 h after induction and transfection with the indicated siRNAs, cells were lysed and subjected to Western blotting. Representative blots are shown, with β-actin as loading control. Graphs are mean values with S.E.M. error bars, based on 6–7 independent replicates per condition. M: mock siRNA, si, s2, s3: three different siRNAs for each MYB. ****p < 0.0001, One-way ANOVA with Dunnet post-test. (D) qPCR analysis of the TIMP2 mRNA level in vector- and p95HER2 cells after knockdown of MYB and MYBL1 (vector cells) or MYBL2 (p95HER2 cells). Reference genes are GAPDH and β-actin. Data are means with S.E.M. error bars, of 3 independent experiments. ***Significantly different from the corresponding mean values in vector cells, p < 0.05 and 0.01, respectively. ^###Significantly different from the corresponding mean values in mock-transfected p95HER2 cells, p < 0.05 and 0.01, respectively. (E) Relative TIMP2 protein level is increased by p95HER2 expression in MCF-7 cells. The graph shows mean values with S.E.M. error bars, of 3 independent experiments. *Significantly different from the value in vector cells, p < 0.05. (F,G) Relative TIMP2 protein level is increased after knockdown of MYB and MYBL1 (vector cells) or MYBL2 (p95HER2 cells). ***Significantly different from the corresponding mean values in mock-transfected cells, p < 0.05 and 0.01, respectively.