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. 2021 May 5;8:625323. doi: 10.3389/fvets.2021.625323

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Copyright © 2021 Kiser, Wang, Zanella, Scraggs, Neupane, Cantrell, Van Tassell, White, Taylor and Neibergs.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) SNP272 is not responsible for bta-miR-1197's targeting of EDN2 3-UTR (P > 0.05). miR-1197 and its predicted seed binding site in the 3′-UTR of EDN2 (top). SNP272 is highlighted in red. (B) bta-miR-2339 does not target 3′-UTR of EDN2 (P > 0.05). miR-2339 and its predicted seed binding site in the 3′-UTR of EDN2 (top). HEK293 cells were co-transfected with EDN2-3′-UTR luciferase constructs (SNP272 A/G) and microRNA precursors (miR-1197 or miR-2339). microRNA negative control oligonucleotides were used as negative controls. Luciferase activities (in triplicates) were measured 24 h after transfection. Renilla luciferase activities were normalized against firefly luciferase activities, and mean normalized Renilla luciferase activities (±SD) from three independent experiments were determined and expressed relative to control values.