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. 2017 Sep 29;7:12401. doi: 10.1038/s41598-017-10861-6

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© The Author(s) 2017

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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Significant polymorphic methylation susceptible loci (MSL) in larvae. (a) Significant loci between groups of larvae subjected at 15 °C, 17 °C or 19 °C from 0 to 15 days post fertilization plotted as a heatmap (n = 57 significant loci). (b) Significant loci between groups of larvae subjected to constant 15 °C or 19 °C, and larvae switched from 15 °C to 19 °C at 15 hours post fertilization (hpf; 15–19(15)), 120 hpf (15–19(120)) or 240 hpf (15–19(240)), or switched from 19 °C to 15 °C at 15 hpf (19-15(15)), 120 hpf (19-15(120)) or 240 hpf (19-15(240)). In this case, the n = 68 significant loci common to both types of temperature switches, (15 °C to 19 °C and 19 °C to 15 °C) were identified and used for the heatmap. Significant single loci were identified after applying multiple Fisher’s exact tests and adjusting the obtained p-values by the method of Benjamini and Hochberg. Only loci with an FDR < 0.05 were used in the analysis. A hierarchical clustering using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) based on pairwise Gower’s distances of categorical variables was applied for the loci. Four types of loci were used: Type I, unmethylated; Type II, inner cytosine methylation; Type III, hemi-methylation of the outer cytosine; Type IV, full methylation.