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. 2019 Nov 26;39(9):1983–1996. doi: 10.1038/s41388-019-1120-z

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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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Fig. 4 — miR-200a overexpression specifically resulted in JNK2 induction, in turn leading to c-Jun phosphorylation and MMP-2 transcription. a–d Cell extracts obtained from the indicated transfectants were subjected to western blot for determination of protein expression. β-Actin or α-tubulin were used as protein loading control. e shJNK2 and Nonsense were stably transfected into T24T(miR-200a) cells and total RNA was extracted; MMP-2 mRNA levels in cells were evaluated using real-time PCR. Bars represent means ± SD from three independent experiments. f Wild-type MMP-2 promoter-driven luciferase reporter was co-transfected together with pRL-TK into T24T(miR-200a/nonsense) and T24T(miR-200a/shJNK2) cells. After 24 h of transfection, transfectants were used to evaluate for luciferase activity. Results presented as MMP-2 promoter activity relative to vector control transfectants. Each bar represents means ± SD from three independent experiments. Invasion abilities of T24T(miR-200a/Nonsense) vs. T24T(miR-200a/shJNK2) cells and UMUC3 (miR-200a/Nonsense) vs. UMUC3 (miR-200a/shJNK2) cells were evaluated using invasion chamber (g, i); relative invasion ability was plotted (h, j). Bars represent means ± SD from three independent experiments. *Significant difference (p < 0.01)