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. 2018 Jul 23;8:11096. doi: 10.1038/s41598-018-29420-8

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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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Validation of RNA-Sequencing results by quantitative RT-PCR. (A) Log₂ fold change comparison of RNA-Seq (dark blue bars) and quantitative real-time PCR (qRT-PCR, light blue bars) for five differentially expressed genes downregulated at D3 (LABLA, CSN2, CSN1S2, CSN1S1, SLC7A5; n = 12), five genes upregulated at D3 (MXRA5, SLC7A8, LBP, ANGPTL4, LOXL4; n = 12), and 3 genes with similar expression (CCL28, IGSF3, ZO;3, n = 12) when comparing D3 vs. D-3 relative to dry-off (D0, ~46 d relative to calving). (B) Correlation between RNA-Seq and RT-PCR gene expression (R² = 0.96, p < 0.0001).