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Description: We validated our microarray data using qPCR. 12S rRNA, a mitochondrial gene product 1, served as our internal control. We confirmed that miR-181c was predominantly localized to the mitochondrial fraction; its expression in the mitochondrial fraction was similar to that in the total heart fraction (Fig. 2A). In addition, fluorescence in situ hybridization (FISH) (Fig. 2C) shows co-localization of miR-181c with the mitochondrial marker, mitoTracker red. We also examined miR-181c expression in both cytosolic and mitochondrial fractions (Fig. 2B, left panel), using qPCR and found that miR-181c was enriched primarily in the mitochondrial fraction. We evaluated several other miRNAs as negative controls for mitochondrial localization, and miR-1192 data are included in Fig. 2B (right panel). We used 5S rRNA as an internal control for these sets of qPCR data, as it is known that this gene is present in both the cytosol and mitochondrial fractions 18. Data are collected from Figure 2.
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