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. 2020 Jul 30;8:669. doi: 10.3389/fcell.2020.00669

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Copyright © 2020 Jankowska, McGill, Pezeshkpoor, Oldenburg, Sauna and Atreya.

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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Predicted miRNAs indeed target 3′UTR of F8-mRNA. (A) Relative luciferase activity in HEK-293T and Hep-G2 cells transfected with luciferase reporter plasmid pEZX-MT05 and miRNA expression vectors (pEZX-MR04), for miR-144, miR-19b and miR-186 and scrambled miRNA as control (SC) (Mean ± SEM for HEK293: SC = 0.993 ± 0.024, N = 15; miR-144 = 0.977 ± 0.017, N = 6; miR-19b = 0.7656 ± 0.017, N = 6; miR-186 = 0.816 ± 0.035, N = 15 and for Hep-G2: SC = 0.999 ± 0.078, N = 6; miR-144 = 0.962 ± 0.013, N = 6; miR-19b = 0.897 ± 0.032, N = 6; miR-186 = 0.782 ± 0.055, N = 9). (B) QPCR analysis to determine overexpression level of miRNAs 72 h after transfection (Mean ± SEM for HEK-293T: SC = 1.020 ± 0.023, N = 4; miR-144 = 946505 ± 40246, N = 3; miR-19b = 320.2 ± 31.61, N = 3; miR-186 = 17.39 ± 1.91, N = 6 and for Hep-G2: SC = 1.001 ± 0.039, N = 4; miR-144 = 5358 ± 208.3, N = 3; miR-19b = 124.5 ± 8.98, N = 3; miR-186 = 4.392 ± 0.257, N = 8) and (C) expression of endogenous miRNAs: miR-144, miR-19b and miR-186 in whole blood (WB from two healthy donors), HEK-293T, liver cell lines: Hep-G2, Huh-7, and HeLa, relative to LCL cells (Mean ± SEM for miR-144: LCL = 1.281 ± 0.287, N = 6; WB = 386,400 ± 84274, N = 6; HEK-293 = 1.100 ± 0.057, N = 3; Hep-G2 = 25.39 ± 2.53, N = 3; Huh-7 = 97.2 ± 6.73, N = 3 and HeLa = 23.94 ± 4.11, N = 3; for miR-19b: LCL = 1.020 ± 0.089, N = 6; WB = 4.303 ± 1.082, N = 6; HEK-293 = 63.34 ± 5.204, N = 3; Hep-G2 = 7.820 ± 1.190, N = 3; Huh-7 = 0.985 ± 0.005, N = 3 and HeLa = 7.225 ± 0.935, N = 3; and for miR-186: LCL = 1.015 ± 0.075, N = 6; WB = 235.9 ± 28.74, N = 6; HEK-293 = 49.02 ± 4.074, N = 3; Hep-G2 = 36.14 ± 1.414, N = 3; Huh-7 = 3.336 ± 0.362, N = 3; and HeLa = 36.03 ± 0.81, N = 3). *P < 0.05; **P < 0.01; ***P < 0.001.