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. 2017 Jun 8;7:3037. doi: 10.1038/s41598-017-02873-z

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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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RNA-seq reproducibility and validation. (a) Growth curves of the five strains (yA635, yA995, yA1382, yA637, yA639) that were subjected to RNA-seq analysis. RNA was isolated before the addition of 0.3 M CaCl₂ (T₀), after 45 minutes of CaCl₂ addition (T₄₅), and after ~9 hours (T_end). Samples were subjected to RNA-seq or northern analyses. (b) Spearman’s correlation heatmap of unclustered RNA-seq biological replicates for all strains at three timepoints. Note that the scale of the Spearman values starts at 0.9. Actual values are provided in Supplementary Table 2. (c) Northern analyses of total RNA extracted from wild-type (yA635) and puf1Δpuf2Δ (yA639) strains for untreated (T₀) and 0.3 M CaCl₂ treated (T₄₅) samples. Radiolabeled PCR products of the genes indicated to the right were used as probes. (d) Bands in the northern blot (c) were quantified with ImageQuant, and the ratio for the signals at T₄₅ to T₀ are presented for each gene (stripes). RNA-seq results (weave) for the same genes from the same biological replica are presented. Compatibility of the results was calculated using Pearson correlation (p value = 0.005).