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. 2018 Jun 25;8:9630. doi: 10.1038/s41598-018-27719-0

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© The Author(s) 2018

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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Identification of target genes of key miRNAs during hepatocyte differentiation. (A) Strategy for predicting common targets of miR-192-3p and miR-192-5p during HD. (B) TaqMan qPCR analysis confirming that miR-192-3p/5p were specifically upregulated during HD (n = 3 independent cultures for each time-point). (C) qPCR results showing the expression tendencies of common targets of miR-192-3p/5p during HD (n = 3 independent cultures for each time-point). (D) Correlation plot revealing reverse-correlations between miR-192-3p/5p and MGAT4C/CALN1/ADCYAP1, respectively. (E) Luciferase reporter assay confirming that miR-192-3p/5p could inhibit the 3’UTR of MGAT4C (n = 3 independent cultures for each group). (F) qPCR results showing the expression of ectodermal marker MGAT4C in HD 6 cells upon transfection of miR-192-3p/5p mimics (n = 3 independent cultures for each group). (G) qPCR results showing the expression of MGATC4C in HD 12 cells upon transfection of miR-192-3p/5p inhibitors (n = 3 independent cultures for each group). In (C), data are presented as the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 for statistical comparisons between day 0 and other time-points (ANOVA plus Bonferroni’s post hoc test). In (E–G), data are presented as the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 for statistical comparisons between control groups and experimental groups (ANOVA plus Bonferroni’s post hoc test). OE: overexpression; KnD: Knockdown; KD: nephron progenitor differentiation; NC: non-targeting control.