Fig. 3

miRNAs/isomiRs showing differential expression on D12, D16 and D20 of the estrous cycle and early pregnancy. a Dendrogram representing results of unsupervised hierarchical clustering (HCL) created with Pearson correlation coefficient. Rows indicate single miRNAs, while columns represent different days of the estrous cycle and pregnancy. b Clusters generated by Self Organizing Tree Algorithm (SOTA) representing expression profiles of mature, canonical miRNAs expressed in the porcine endometrium of pregnant and cyclic pigs. c Venn diagrams demonstrating relations between DE miRNAs and isomiRs for selected comparisons during pregnancy. Marked up- and down-regulated miRNAs were chosen for microarray validation by real-time RT-PCR. d Relative expression of DE miRNAs selected for validation of Illumina sequencing, using stem-loop real-time RT-PCR. miRNA expression on different days of the estrous cycle and pregnancy (n = 7-9 animals/group) is presented relative to the reference let-7d-3p. Two-way ANOVA analysis with Bonferoni post-hoc test was performed to find statistical differences. Results are presented as boxplots with means ± SEM. Means with different superscripts differ significantly (small letters – the estrous cycle, capital letters – pregnancy). * p < 0.05, indicate difference between reproductive states (estrous cycle vs. pregnancy) within the same day. AU – arbitrary units