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. 2020 Nov 13;3:677. doi: 10.1038/s42003-020-01404-3

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© The Author(s) 2020

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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Fig. 7 — a, b Female nude mice are i.p. injected with SKOV3, OVCAR-3, A2780, or A2780 CIS cells and treated with vehicle (Ctr) or MAC (30 mg/kg oral daily) for 5 weeks. Graph represents the number of visible metastasis. Columns show the mean ± SD (*p < 0.001 vs. Ctr). c WB analysis of the indicated proteins in lysates from i.p. nodules of three representative (M1-M3) SKOV3, OVCAR-3, A2780, and A2780 CIS xenografts treated as in a. β-actin is used as loading control. d ZEB1, Vimentin, and E-cadherin gene expression in i.p. nodules of SKOV3, OVCAR-3, A2780, and A2780 CIS xenografts treated as in a is analyzed by qRT-PCR and normalized to cyclophilin-A. Values are the mean ± SD (*p < 0.004 vs. Ctr). e miR-200b/c expression in i.p. nodules of SKOV3, OVCAR-3, A2780, and A2780 CIS xenografts treated as in a is analyzed by qRT-PCR. U6 is used to normalize. Values are the mean ± SD (*p < 0.006 vs. Ctr). f The binding of ZEB1 on miR-200b promoter region and on a region -1900 bp upstream the miR-200b TSS site (lacking ZEB1 binding sites, negative control for non-specific enrichment) is analyzed by ChIP assay followed by PCR in SKOV3 xenografts treated as in a. Anti-IgG mouse (IgGM) Ab is used as control for all ChIP reactions. g Schematic model of a regulatory circuit established by ET_AR-miR-200b/c-ZEB1 to control metastatic progression that is activated by ET-1 and inhibited by macitentan.