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. 2018 Feb 27;8:3704. doi: 10.1038/s41598-018-21735-w

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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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miR-214 expression correlates with skeletal elements in zebrafish. (A) Expression of miR-214 during zebrafish development, determined by miRNA qPCR. Values were normalized using zebrafish U6 small RNA and 24 hpf as reference sample and represent the mean ± s.d. of at least 3 independent replicates. hpf hours post fertilization, dpf days post fertilization, N.D. non-detected. Gap in the y-axis separates two different scales. (B) Detection of miR-214 by in situ hybridization in zebrafish with 10 (a), 20 (b, c, d) and 90 (e, f, g, h, i) dpf. From head to tail, miR-214 was detected in eye lens (arrowhead, a, b), retina (white arrowheads, a, b), brain (arrow, a), chondrocranium (asterisk, a, b), pharyngeal cartilage (white arrows, a, c), kidney (arrows, d), scales (arrow, e), muscle myotomes (arrowheads, e), cartilage in the base of pectoral fins (arrows, f), notochordal sheath (arrow, g), osteoid of haemal arches (arrows, h) and growth zones of vertebral body (arrowheads, i). Hybridization with negative control (scrambled) probe did not produce detectable signal, as observed in 90 and 20 dpf specimens (j and k, respectively). Scale bars: 0.2 mm for a, e, f and k; 0.1 mm for b, c, d, h, i and j; and 0.05 mm for g. (C) Relative expression of miR-214 in zebrafish (left panel) and mouse (right panel) adult tissues, determined by miRNA qPCR. Values were normalized using U6 small RNA and muscle as reference sample and represent the mean ± s.d. of at least 3 independent replicates (one-way Anova, different letters indicate statistical significance, p < 0.05). B. arches, Branchial arches. dpf days post fertilization.