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. 2019 Jul 5;9:9776. doi: 10.1038/s41598-019-46170-3

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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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Effects of miR-214 overexpression on prostate cancer cell migration and invasion. (A) Wound healing assay: RWPE-2, PC3, DU145, and MDA-PCa-2b cells were transfected with NC or miR-214 mimic for 48 h. The effect of the miR-214 expression on cell migration was analyzed using the wound healing assay. Representative images of the wound healing assay (top) and the calculated scratch area (bottom) are shown. (B) 3D spheroid invasion assay: RWPE-2, PC3, DU145, and MDA-PCa-2b cells were transfected with NC or miR-214 mimic and then analyzed using the 3D spheroid assay. 3D spheroid images (top) and the quantified 3D invasion area (bottom) were determined by subtracting pre-invasion (day 3) from post-invasion (day 10) area. (C) Trans-well invasion assay: PC3 cells were transfected with NC or miR-214 mimic. After 48 h, the effect of miR-214 mimic on cell invasion was analyzed by trans-well invasion assay in miR-214-transfected PC3 cells stained with Calcein-AM (Green fluorescence, top) and quantified (bottom). The invasion assay was performed in duplicate, and results are representative of three independent experiments. (D,E) The effects of NC or miR-214 mimic transfection on the expression of E-Cadherin (top, green) and N-Cadherin (bottom, green) in PC3 cells were analyzed by (D) immunofluorescence and (E) western blotting. *P < 0.05, **P < 0.005, ***P < 0.0005 compared with NC-transfected cells.