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. 2020 May 25;12(2):107–127. doi: 10.1007/s13238-020-00723-9

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Loss of 4.1N promotes EOC cells EMT in vitro. (A) Representative images of transwell migration assay (top) and transwell invasion assay (bottom) of A2780 cells and SKOV3 cells with 4.1N-expression and 4.1N-loss. (B) Graphical representation of transwell migration assay and transwell invasion assay. The transwell migration assay of A2780 OE cells (31.25 ± 6.11) showing reduction in transwell migration, compared to empty vector control (215.63 ± 27.44) (n = 8, 2-tailed unpaired t-test, P < 0.001). The transwell migration assay of SKOV3 KO cells (11.13 ± 2.17) showing increase in transwell migration, compared to empty vector control (8.75 ± 1.83) (n = 8, 2-tailed unpaired t-test, P = 0.033). The transwell invasion assay of A2780 OE cells (18.63 ± 1.77) showing reduction in transwell invasion, compared to empty vector control (43.63 ± 6.07) (n = 8, 2-tailed unpaired t-test, P < 0.001). The transwell invasion assay of SKOV3 KO cells (7.50 ± 1.85) showing increase in transwell invasion, compared to empty vector control (3.50 ± 0.76) (n = 8, 2-tailed unpaired t-test, P < 0.001). All these means ± SD results from three independent experiments. Scale bar, 50 µm. (C) Representative graph of the invasion curve using real-time cell analyzer (RTCA). The rate of invasion was monitored in real-time using the xCELLigence system (n = 4). (D) Representative images of scratch wound healing assay of A2780 cells and SKOV3 cells with expression of 4.1N and respective control. (E) Graph of scratch wound healing assay of A2780 cells and SKOV3 cells with 4.1N-expression and 4.1N-loss. A2780 OE cells showing 37.7% reduced wound area, compared to control wound area (n = 3, 2-tailed unpaired t-test, P < 0.05); compared to SKOV3 KO cells, SKOV3 KO cells indicate 61.5% increase (n = 3, 2-tailed unpaired t-test, P < 0.05). (F) Effects of 4.1N on EMT-related mRNA expression was assayed by qRT-PCR and results means ± SD from three independent experiments. β-Actin was used as internal control. (G) Effects of 4.1N on EMT-related proteins expression was assayed by Western blot. β-Actin was loading control. *P < 0.05, **P < 0.01, ***P < 0.001