miRNA detection frequency.

<p>Most highly detected miRNAs grouped on mature sequence, normalised to total annotated reads (expressed in reads per million mapped (RPMM) from 3 rat mid-myocardial samples).</p

Novel rat miRNA similar to miR-676-3p.

<p>A: Alignment of the novel rat miRNA sequence to miR-676-3p sequences as published in miRBase v18. B: Predicted stem-loop structure (mFold 3.2) of proposed pre-miR deduced from genomic sequence. Mature miRNA highlighted in blue.</p

Relative miRNA abundance by TaqMan assay.

<p>Expression of miRNAs relative to rno-miR-1 according to 2^−ΔCt where ΔCt = Ct – Ct_miR-1 (mean ± SEM). Ct values were compared by repeated measures ANOVA and Tukey’s multiple comparison test; **p<.01, ***p<.001. Mid-myocardial samples from n = 6 hearts.</p

Length distribution analysis of small RNA sequences in 3 mid-myocardial rat samples.

<p>Reads greater than 12 nt are included. Red bars represent reads annotated to known rat miRNAs (miRBase v18), green bars represent reads annotated to miRNAs in other species (novel orthologs) and black bars represent unannotated small RNA sequences.</p

The rat miR-486 gene.

<p>A: Alignment of the genomic miR-486 sequence across nine species including rat (highlighted in red). Blue shading indicates percentage sequence identity and the mature and ‘star’ sequences represented by the most common reads are indicated with arrows. B: Predicted stem-loop structure (mFold 3.2) of pre-miR-486 deduced from genomic sequence. Mature miRNA highlighted in blue.</p

Frequency of isomiRs with 5′ and/or 3′ variations.

<p>Distributions of 5′ (left) and 3′ (right) end variants are shown for miRNAs derived from the 5′ arm (A) and from the 3′ arm (B) of the pre-miR hairpin. Data are mean ± SEM for three mid-myocardial samples.</p

Epicardial/endocardial differential expression of unique sequences (≥ ±2 fold difference for miRs of ≥100 mean normalised reads detected in either layer). P<0.05 Baggerleys multiple comparison test.

<p>Epicardial/endocardial differential expression of unique sequences (≥ ±2 fold difference for miRs of ≥100 mean normalised reads detected in either layer). P<0.05 Baggerleys multiple comparison test.</p

Epicardial/endocardial differential expression of exact mature (or mature*) miRNAs (≥ ±1.5 fold difference for miRs of ≥10 raw reads detected in either layer).

<p>P<0.001,</p><p>P<0.01, Baggerleys multiple comparison test.</p

Differential suppression of a gelsolin sequence-tagged reporter gene by miR-133a isomiRs.

<p>HEK293 cells were transfected with expression plasmids encoding mCherry with a 3′ partial gelsolin sequence and pSM30 containing inserts for miR-133a, miR-133a(v), a random non-targeting sequence (NTC) or siRNA against mCherry (siR-mCh). Cells were imaged and analysed for both green and red fluorescence intensity. A decrease in red/green ratio <i>vs</i> NTC indicates downregulation of target gene expression. Data were log transformed and compared by one-way ANOVA with Bonferroni's Multiple Comparison Test. All pairwise comparisons were significant (p<.001). Comparison of miR-133a <i>vs</i> miR-133a(v) is indicated (***). Number of cells (n): miR-133a 4640; miR-133a(v) 2843; NTC 4088; siR-mCh 3884. Representative of 3 separate experiments in which miR-133a(v) was significantly more effective than miR-133a.</p

Pathway analysis of MAP Kinase signaling (Kegg pathway).

<p>Highlighted genes are targets predicted by 2 or more algorithms of the top 16 highest detected miRNAs (boxed in red; P = 1.86×10⁻⁶; 75 targets) showing overlap with genes expressed in the heart (highlighted; expression data 9 SD rat left ventricular myocardium samples from 2 microarray experiments, (GSE6943; GSM160095–100 & GSE6880; GSM158589–91; <a href="http://www.ncbi.nlm.nih.gov/gds" target="_blank">http://www.ncbi.nlm.nih.gov/gds</a>).</p