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. 2021 Dec 22;41(8):1178–1189. doi: 10.1038/s41388-021-02084-x

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© The Author(s) 2021

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 Schematic representation of predicted miR-138 MREs on the MYC transcript. The thick line indicates the MYC CDS, while the thin lines indicate the UTRs. B Effect of miR-138 overexpression on the luciferase activity of the respective MRE-reporter constructs in HCT116 (left panel) and DLD-1 (right panel) cell lines. RC denotes the reverse complement of miR-138 which was used as positive control. C Schematic representation of the ATG-less MYC CDS cloned into the psiCHECK-2 vector. Location and sequences of the wild-type (WT) and mutant MREs A (yellow oval) and C (green oval) are depicted. Red font indicates the mutated nucleotides. The cross indicates the absence of the start codon ATG. D Effect of miR-138 overexpression on the luciferase activity of the MYC CDS reporter constructs with mutated MREs (CDS mut) compared to the wild-type CDS reporter (CDS WT) in HCT116 (left panel) and DLD-1 (right panel) cell lines. E Effect of miR-138 overexpression on the luciferase activity of the Renilla luciferase (RLuc) gene fused with MRE A in HCT116 (left panel) and DLD-1 (right panel) cell lines. Top panel illustrates the modification of RLuc gene construct. F Effect of miR-138 overexpression on the luciferase activity of the RLuc and MYC CDS fusion reporter construct with mutated MRE A (mut_A) compared to the wild-type MYC CDS (WT) in the HCT116 (left panel) and DLD-1 (right panel) cell lines. The top panel illustrates the fusion gene construct. G RT-qPCR analysis of MYC transcript enrichment upon biotinylated miR-138 pulldown in HCT116 (left panel) and DLD-1 (right panel) cell lines. (B, D, E and F) Mean ± SEM; N ≥ 3. G Mean ± STD; N ≥ 3. *P < 0.05; **P < 0.01; ***P < 0.001.