Figure 8

Search Tool for the Retrieval of Interacting Genes/Proteins (STRING)—based analysis uncovered possible mediators of miR-378 action in the liver. The STRING analysis was performed on differentially expressed genes that were oppositely regulated in mdx vs. WT and mdx/miR-378^−/− vs. mdx livers. (A) 7 out of 20 genes that were downregulated in mdx vs. WT and upregulated in mdx/miR-378^−/− vs. mdx mice were found by STRING to interact with each other and exhibit co-expression patterns. Five of them indicated by asterisks were predicted by miRmap database to be direct targets of miR-378. (B) Verification of RNA sequencing results by qRT-PCR analysis of ATP citrate synthase (Acly), fatty acid synthase (Fasn), glycerol-3-phosphate acyltransferase, mitochondrial (Gpam), patatin-like phospholipase domain-containing protein 3 (Pnpla3), and stearoyl-CoA desaturase-1 (Scd1) predicted as miR-378 targets along with perilipin 2 (Plin2) and glycerol-3-phosphate acyltransferase 3 (Agpat9); n = 4–8/group, together with (C) gene ontology (GO) terms indicating pathways affected by interacting genes. (D) The expression of 2 out of 22 genes, hemolytic complement (Hc) and complement factor properdin (Cfp), that were higher in mdx vs. WT and lower in mdx/miR-378^−/− vs. mdx liver based on RNA-seq data, was verified by qRT-PCR analysis; n = 4–8/group. (E) Those genes, presented together with the affected GO terms, were shown by STRING to interact with each other. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001 by one-way ANOVA with Tukey’s post-hoc test; ^#p < 0.05 additionally tested by Student’s t-test for comparison of mdx vs. WT and mdx/miR-378^−/− vs. mdx only.