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. 2017 Jun 15;7:3568. doi: 10.1038/s41598-017-03362-z

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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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EZH2 regulates TIMP2 expression in vitro. (a) Western blot analysis of EZH2, TIMP2, MMP2, and MMP9 protein expression in ovarian cancer cell lines. Histone-3 (H3) and β-actin serve as internal controls. The cropped blots are used in the figure, and full-length blots are presented in Supplementary Fig. S5. (b) EZH2 and TIMP2 mRNA expression levels in seven ovarian cancer cell lines were detected by quantitative real-time PCR. (c) Negative correlation between TIMP2 and EZH2 mRNA levels in ovarian cancer cell lines. (d) qRT-PCR for EZH2 and TIMP2 expression in A2780 and SKOV3 cells with EZH2 up- or down-regulation by transfection with an EZH2-overexpressing plasmid (EZH2) or targeting shRNA (shEZH2). (e) Western blot analysis of EZH2, TIMP2 and MMP2/9 protein expression in EZH2-regulated A2780 and SKOV3 cells. Histone-3 (H3) and β-actin are used as internal controls. The cropped blots are used in the figure, and full-length blots are presented in Supplementary Fig. S6. (f) Immunocytochemical staining of EZH2 and TIMP2 in EZH2-regulated A2780 and SKOV3 cells (scale bar, 20 μm). (g) Gelatin zymography assays of MMP2 and MMP9 activity in EZH2-regulated A2780 and SKOV3 cells. The cropped gels are used in the figure, and full-length gels are presented in Supplementary Fig. S7. *P < 0.05; **P < 0.01; ***P < 0.001.