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. 2019 Jun 10;9:8418. doi: 10.1038/s41598-019-44828-6

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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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ELK3 determines the miRNA contents of exosomes derived from LECs. (A) miRNA expression comparison between siNS LECs and siELK3 LECs. (Left) Scatter plot of the miRNA microarray analysis. The axes of scatter plot are normalized data from the samples. (Right) Volcano plot representing the p-values (Y axis) for the observed differences in miRNA expression (X axis). Three representative upregulated miRNAs were labeled in blue, and 3 representative downregulated miRNAs were labeled in red. (B) The analyzed representative miRNAs, fold changes from the miRNA microarray results and miRNA status as a tumor suppressor miRNA or onco-miRNA. (C) The intracellular and exosomal levels of 3 miRNAs that were selected as candidate antioncogenic miRNAs were analyzed by qRT-PCR. (D) Transwell assay of MDA-MB-231 cells that were transfected with the indicated miRNA mimics (Upper). The expression level of each miRNA was analyzed by qRT-PCR (Lower). Scale bar represents 100 μm. (E) The intracellular and exosomal levels of 3 miRNAs selected as candidate pro-oncogenic miRNAs were analyzed by qRT-PCR. (F) The effect of miR-30e-3p mimic or miR-30e-3p inhibitor on the proliferation of MDA-MB-231. The expression level of miR-30e-3p for each treatment was analyzed by qRT-PCR and the effect of each miRNA on the cell proliferation was assessed by cell counting at the indicated time (Left). Cell morphology was visualized via light microscopy at 48 h after transfection of each miRNAs (Right). Scale bar represents 200 μm. (G) Transwell assay of MDA-MB-231 cells that were transfected with the miR-30e-3p mimic or miR-30e-3p inhibitor. Scale bar represents 100 μm. (H) Kaplan-Meier plots for miR-503, miR-4269 and miR-30e. The plots for miR-503 and miR-30e were analyzed with all subtypes of breast cancer patients from the METABRIC dataset, and the plot for miR-4269 was analyzed with triple-negative breast cancer patients from the METABRIC dataset. Error bars represent the standard error from three independent experiments, and each experiment was performed using triplicate samples. *P < 0.05, **P < 0.01 ***P < 0.001 and ****P < 0.0001 (Student’s t-test).