miRNA profiling in heart tissue of AGAT-/- mice

miRNA sequencing was performed in left ventricular heart tissue of wt (n = 5), AGAT-/- (n = 5), and AGAT-/- mice supplemented with homoarginine (n = 5). Small RNA libraries were prepared using TruSeq Small RNA Sample Preparation Kits (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. The CLC Genomics Workbench (clcbio.com/products/clc-genomics-workbench/) was used to map reads from miRNA sequencing against the murine set of all known miRNAs, which was retrieved from miRBase (www.mirbase.org/). The number of reads falling in mature miRNAs were extracted and further processed in R. Only miRNAs covered by more than ten reads were kept for further analyses. Differential expression of miRNAs between groups of mice was calculated by R/Bioconductor package DESeq2 and the False Discovery Rate (FDR) based Benjamini-Hochberg method was used to account for multiple tests. Differentially expressed miRNAs with an FDR = 0.05 were considered significant. Overall design: Heart tissue miRNA profiles in wt, AGAT-/- and AGAT-/- mice supplemented with homoarginine were generated by deep sequencing using Illumina HiSeq 2500.