Molecular Network Approach Reveals Rictor as a Central Target of Cardiac ProtectomiRs

Figures

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  Figure 1. Experimental protocol.

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  Figure 2. Predicted interaction network of ProtectomiR microRNAs (A) showing the network neighborhood of predicted hub genes with at least three microRNA–mRNA target interactions (node degree) (B,C). Panel (C) shows the role of the most central target hub Rictor. MicroRNAs upregulated and downregulated in pre- and/or postconditioning and target mRNAs are indicated in green, red and blue, respectively. Dark blue nodes represent mRNAs with a node degree of at least 3. A high-resolution version of the network is available as Supplementary Figure S1.

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  Figure 3. Pig homologues of the Rictor targeting microRNAs (A) were identified based on rat–pig microRNA sequence similarity. Four of the five rat microRNAs (miR-139-5p, miR-320, miR-212, miR-503) showed a 100% match and one (rno-miR-188-5p vs miR-362) showed a 56% match between pig and rat microRNA homologues (B). Expression levels of the five Rictor targeting ProtectomiR microRNA homologues were tested with qPCR in ischemic preconditioning and postconditioning compared to ischemia-reperfusion (C). Black circle, triangle and square signs represent individual data points. * p < 0.05 vs. Isch group, n = 4–6, one-way ANOVA followed by Dunnett post-hoc test.

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  Figure 4. Central target hub, Rictor mRNA expression was measured with qPCR in the ischemic zone (A) and in the non-infarcted remote zone (B) of the porcine left ventricular myocardium. RICTOR protein expression was measured with Western blot in the ischemic zone (C) and in the non-infarcted remote zone (D) of the porcine left ventricular myocardium and normalized to total protein staining. Black circle, triangle and square signs represent individual data points. * p < 0.05 vs. Isch group, n = 4–6, one-way ANOVA followed by Dunnett post-hoc test.

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  Figure 5. Gene ontology enrichment analysis (biological processes) of all microRNA target mRNAs (n = 882) highlights the effect of the ProtectomiR microRNAs on glucose homeostasis, skeletal muscle development, angiogenesis and membrane transport. A total of 21 biological processes out of the 25 with the highest fold enrichment value are presented here. * p < 0.05, vs. control (gene ontology enrichment analysis with Bonferroni correction).