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. 2021 Dec 21;13(1):25. doi: 10.1038/s41419-021-04470-5

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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. 3 — a Predicted base-pairing between miR-708 and Meg3 in human and mouse. Mutants (MUT) of miR-708 and Meg3 were generated by site-specific mutation of the paired bases. b DLD1 and RKO cell lysates were incubated with biotin-labeled Meg3 (WT and MUT types). RT-qPCR analyses were performed to detect miR-708 levels in the RNA pulldown obtained using biotin-labeled Meg3. c DLD1 and RKO cell lysates were incubated with biotin-labeled miR-708 (WT and MUT types). RT-qPCR analyses were performed to detect Meg3 levels in the RNA pulldown obtained using biotin-labeled miR-708. d Luciferase activities were measured in DLD1 and RKO cells. The cells were transfected with the indicated plasmids. e An AGO-RIP assay was used to measure Meg3 levels following miR-708 knockdown in DLD1 cells (left) or overexpression in RKO cells (right). f The relative levels of Meg3 were detected by qPCR in DLD1 and RKO cells constitutively expressing miR-708 or in DLD1 and RKO cells transfected with miR-708 antagomir. g RT-qPCR analyses were performed to detect the relative levels of Meg3 in colonic cancer tissues from AOM/DSS-induced WT (+/+) and miR-708 KO (−/−) mice or from Apc^min WT (+/+) mice and Apc^min miR-708 KO (−/−) mice. h RT-qPCR analyses were performed to detect the relative levels of Meg3 in organoids from Apc^min WT (+/+) mice or Apc^min miR708 KO (−/−) mice. Organoids were treated with miR-708 agomir or miR-708 antagomir and cultured for 6 days. i Representative images of organoids, derived from Apc^min mice, after 6 days of culturing (scale bar: 100 μm). Meg3 interference adenovirus (Adv-Meg3i) or control adenovirus was administered to organoids that had been transfected with miR-708 antagomir or control antagomir. j Representative images of organoids, derived from Apc^min mice, after 8 days of culturing (scale bar: 100 μm). Meg3 overexpression adenovirus (Adv-Meg3) or control adenovirus was administered to organoids that had been transfected with miR-708 agomir or control agomir. The results are presented as the mean ± S.D. in panels (b–j). NS not significant (non-paired two-tailed Student’s t test).