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. 2020 Feb 13;11(2):123. doi: 10.1038/s41419-020-2293-7

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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. 2 — a Dual luciferase reporter gene analysis showed the direct binding between miR-122 and the 3′UTR wild type of CPEB1. The miRNA negative control mimics and the 3′UTR mutant type of CPEB1 have lost their binding capacity for each other. b RIP result showed circRNA_002581 in NIH3T3 cell lysis was pulled down and detected by qPCR with selected primer, indicating the existence of binding between circRNA_002581 and miR-122, where miR-218 was used as negative miRNA control and GAPDH was applied as control. c RNA FISH showed the co-localization of miR-122 (green) and circRNA_002581 (red) in the cytoplasm of HEK-293T cells. The blue spot is the nucleus dyed by DAPI (scale bar: 100 μm). d Western blot showed CPEB1 level was decreased when transfecting HEK-293T cells with miR-122 mimics than with NC mimics. Further co-transfecting circRNA_002581 could antagonize the inhibitory effect of miR-122 mimics on CPEB1 protein level (upper panel); CPEB1 protein level was significantly downregulated when transfected with siCircRNA_002581 in NCTC-1469 cells but significantly upregulated when transfected with miR-122 inhibitor (middle and lower panels). Error bars represent the SD. **p < 0.01.