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. 2018 Oct 23;8:15605. doi: 10.1038/s41598-018-33807-y

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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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(A) The resultant DE miRNAs were analyzed through the use of IPA (Ingenuity Pathway Analysis, QIAGEN Inc., https://www.qiagenbioinformatics.com/products/ingenuitypathway-analysis)¹⁹ to examine the crosstalk between the TGFB1 signaling pathway and miR-34a and miR-16a-5p (seed of miR-424-5p). We revealed a number of genes and pathways, most notably VEGFA, that are reciprocally regulated by these negatively correlated transcription regulators. (B) Total mRNA was purified from CCs denuded from GV COC and M2 COC aspirated during IVF procedures. The mRNAs were subjected to qPCR in duplicate with the examined genes and ACTB primers. Gene expression was calculated relative to the ACTB level in the same sample and expression levels were compared using Student’s t-test. The difference reached p = 0.006 for FOXM1, p = 0.01 for TOP2A and p = 0.08 for CD47. RNAseq (green) and qPCR (blue) results are presented as log2-fold change between COC M2 and COC GV samples.