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. 2019 Sep 27;10:4397. doi: 10.1038/s41467-019-12292-5

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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 — Systematic identification of miR-574-5p/-3p targets in GC cells. a Schematic representation of biotinylated miRNA pull-down experiment. b Classification of miR-574-5p/-3p enriched targets. c, d The expression change of miR-574-5p/-3p pulled down targets versus all genes or miR-574-5p/-3p targets predicted by miRanda in miR-574-5p/-3p overexpressed cells. ER: enrichment ratio. e qRT-PCR validation of the enrichment of the potential targets in miRNA pulled down experiment. f Schematic representation of artificial miR-574-5p/-3p sponges expressing RNAs with six binding sites (6× target) that pair extensively with the seed and 3′-end of the miRNAs but contain a central bulge. g Relative expression of miR-574-5p/-3p in GC cells with overexpression of artificial miR-574-5p/-3p sponges. h Relative luciferase activity of wild type QKI6 or ACVR1B 3′UTRs constructs in 293T and GC cells simultaneously treated with artificial miR-574-5p/-3p sponges. i The protein level of QKI6 or ACVR1B in GC cells with overexpression of artificial miR-574-5p/-3p sponges. Three technical replicates from a single experiment representative of two independent experiments. Data are shown as means ± s.d. **p < 0.01, ***p < 0.001, Student’s t-test.