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. 2020 Jan 16;11(1):34. doi: 10.1038/s41419-019-2188-7

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© The Author(s) 2020

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. 3 — a The binding sites for miR-26a-5p and miR-26b-5p within IL-6 3′-UTR sequence was predicted by starBase. The red nucleotides represent the mutant binding site designed for luciferase reporter assay. b qRT-PCR evaluated IL-6 mRNA levels in HCC cell lines and in normal cells. c ELISA evaluated IL-6 protein levels in HCC cell lines and in normal cells. d, e The effects of DLGAP1-AS1 knockdown in Hep G2 cells and DLGAP1-AS1 overexpression in SNU-387 cells on IL-6 mRNA and protein levels were respectively exhibited using qRT-PCR and ELISA. f RIP assay was performed using the Ago2 antibody to demonstrate the enrichment of DLGAP1-AS1, miR-26a/b-5p and IL-6 mRNA in HCC cells. g RNA pull-down assay was performed to detect the binding ability of IL-6 mRNA with miR-26a/b-5p. h Luciferase reporter assay of IL-6-3′-UTR-WT or IL-6-3′-UTR-Mut reporters elucidated the interaction between IL-6 mRNA and miR-26a/b-5p, and the competing effect of DLGAP1-AS1 to interact with miR-26a/b-5p. All data are presented as the mean ± SD of three independent experiments. **p < 0.01, ***p < 0.001.