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. 2019 Dec 10;8(12):73. doi: 10.1038/s41389-019-0182-7

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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. 3 — a Upper: immunoprecipitation (IP) of Ago2 protein from extracts of HEK-293T cells. Lower: levels of MACC1-AS1 co-precipitated with Ago2 protein were analyzed by RT-PCR. b Upper: schematic of a luciferase reporter in which the entire MACC1-AS1 sequence was fused into the 3′ region of the Renilla luciferase gene of the psiCHECK-2 construct (denoted pLuc-MACC1-AS1). Lower: HEK-293T cells were transfected with pLuc (control) or pLuc-MACC1-AS1, or pLuc-MACC1-AS1 in combination with either the MACC1-AS1 expressing vector or siRNA against MACC1-AS1. Luciferase activities were determined after 36 h transfection using a dual-luciferase assay system. Renilla luciferase activity was normalized to the activity of firefly luciferase. *P < 0.05 as determined by Student’s t test. c Schematic representation of the potential binding sites for individual miRNAs within MACC1-AS1 RNA. d Upper: RT-PCR and agarose gel electrophoresis indicate that MACC1-AS1-MS2 was pulled down by MBP-MCP-conjugated amylose resin. Lower: individual miRNAs were detected by RT-qPCR in the precipitates of MACC1-AS1. **P < 0.01 as determined by Student’s t test. e HEK-293T cells were co-transfected with pLuc, wild-type or mutant MACC1-AS1 reporters with the corresponding miRNA mimics. Luciferase activity was determined by the dual luciferase reporter system. Activity of Renilla luciferase was normalized to the activity of firefly luciferase. **P < 0.01, *P < 0.05 as determined by one-way ANOVA followed by Tukey’s multiple comparison tests.