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. 2022 Jul 13;13:4061. doi: 10.1038/s41467-022-31574-z

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© The Author(s) 2022

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. 5 — a Schematic view of the complex structure generated by the predicted base-pairing interaction between HHIP-AS1 (red) and the 5'UTR of DYNC1I2 mRNA (green). The sequence of hsa-miR-425-5p is also shown and the inferred binding site of this miRNA on DYNC1I2 mRNA 5'UTR is depicted in orange. The scheme highlights how this miRNA-mRNA interaction is predicted to be blocked by HHIP-AS1 association to DYNC1I2 mRNA. Black frame indicates position for zoom out image on the right side. b The wild type 5'UTR of DYNC1I2 mRNA (5'WT) or a mutated version (5'MUT) were cloned in front of a luciferase coding sequence and co-transfected in combination with hsa-miR-425-5p mimics (mir-425) or negative controls (NC) into Daoy cells. Bar graphs indicate the measured relative luciferase activity for each combination ± SEM in three independent experiments. Student’s one-sided t-test; *p < 0.05. n.s. not significant. c DYNC1I2 expression level was measured via qRT-PCR in Daoy, CHLA-266 and HHU-ATRT1 cells upon stable HHIP-AS1 knockdown using two independent stable shRNAs (sh-HHIP-AS1#1 and sh-HHIP-AS1#2), in combination with or without transient inhibition of hsa-miR-425-5p. Bar graphs are presented as the mean ± SD of three independent experiments. d Proliferation rate of Daoy and CHLA-266 with stable knockdown of HHIP-AS1 (sh-HHIP-AS1#1) measured by EdU incorporation in combination with or without transient inhibition of hsa-miR-425-5p. Bar graphs are presented as the mean ± SEM of at least ten independent experiments and corresponding controls were set to 100%. Student’s two-sided t-test; ***p < 0.001; **p < 0.01; *p < 0.05; n.s. not significant. Source data and exact p-values are provided as a “Source Data file”.