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. 2018 Jan 25;8:1570. doi: 10.1038/s41598-018-19817-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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Temporal gene expression of zebrafish ache during embryonic and larval development. (a) Relative ache expression was quantified by qPCR at selected embryonic stages in AB line (N = 1 experiment, n = 20 embryos per group, standard deviation comes from technical replicates). qPCR reactions were run in duplicates and ∆∆Ct was calculated relative to 48 hpf values and normalized with actb2 expression (b) Relative ache expression was quantified by qPCR at 1–5 dpf (N = 2 experiments, n = 20 embryo/larvae per group). qPCR reactions were run in duplicates and ∆∆Ct was calculated relative to 24 hpf values and normalized with actb2 expression. (c) Normalized O.D. 412 values for groups of embryos. AChE enzymatic activity was almost absent in tail-test selected ache−/− mutant embryos when compared with wild-type ache+/? siblings at 28 °C (P = 0.034) or at 33°C (P = 0.003) (performed in triplicates, n = 8 embryos per group). Error bars show the standard deviation.