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. 2021 Jan 4;53(1):52–66. doi: 10.1038/s12276-020-00530-6

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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. 2 — a The effect of cyclopamine, Gant61 or recombinant Shh 10.1038/s12276-020-00530-6 on regulating migration and invasion of JAR cells. Scale bars, 200 μm. b Migrated or invaded cells from panel a were quantified by Image J software. c The effect of cyclopamine, Gant61 orrecombinant Shh on regulating motility of extravillous trophoblasts in first-trimester villous. Scale bars, 500 μm. d Outgrowth area from panel c was quantified by Image J software. e After with or without cyclopamine treatment, expression of p-AKT S473, p-AKT T308, and pan-AKT proteins were detected by western blot. f After with or without cyclopamine treatment, expression of E-Cadherin, VEGF-A, and GAPDH proteins were detected by western blot. g VEGF-Astaining in control group and recurrent miscarriage group were detected by immunohistochemistry. Scale bars, 100 μm. h Expression of VEGF-A from panel g was analyzed and quantified by Image J software. i CD31 staining in control group and recurrent miscarriage group were detected by immunohistochemistry. Scale bars, 100 μm. j Expression of CD31 from panel i was analyzed and quantified by Image J software. *p < 0.05, **p < 0.01, ***p < 0.001.