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. 2018 Jun 22;8:9575. doi: 10.1038/s41598-018-27810-6

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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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Knockdown of Twist in MCF-7 cells significantly inhibits E2-induced production of CCL2. (a) The protein levels of Twist at the indicated time points in MCF-7 cells cultured with 10 nM E2 were determined by Western blot. Cropped images are from samples run on the same gel. Full-length blots are displayed in Supplementary Fig. S4b; (b) The relative levels of Twist expression in MCF-7 cells after siRNA transfection were measured by qRT-PCR and Western blot. Full-length blots are demonstrated in Supplementary Fig. S4c; (c) The levels of E2-induced CCL2 secretion in MCF-7 cells at the indicated time points after knockdown of Twist were detected by ELISA analysis; (d) The mRNA levels of E2-induced CCL2 in MCF-7 cells after knockdown of Twist was determined by qRT-PCR. Data are shown as mean ± SEM and images are representative from three independent experiments. ***P < 0.001 vs. the controls; ns, not significant.