Fig. 5

miR-574-5p exposure in mouse splenocytes induces significant alterations in the expression of genes associated with immune and inflammatory responses. Splenocytes from 8-wk old female B6.WT mice (n = 3) were divided into two equivalent parts, one for 10 µg/ml of Dotap-PS-miR-574-5p and one for Dotap only transfection for 24 h. Gene expression was profiled by RNA-Seq as described on Supplementary Methods. (a) RT-qPCR confirmation of RNA-seq targets in miR-574-5p-transfected mouse splenocytes. qPCR analyses were performed with 28 selected mRNAs (which included 4 down-regulated genes and 24 up-regulated genes as determined by the RNA-Seq) in Dotap-PS-miR-574-5p treated and Dotap-only treated splenocytes as described. (b) 30 top signaling pathways that were significantly enriched for genes that were up-regulated by at least 2-fold as a consequence of miR-574-5p exposure (Dotap-PS-miR-574-5p versus Dotap-only, n = 3) as analyzed by the Reactome pathway analysis. The digital number next to each bar represented the count of the significantly-up-regulated genes enriched in the pathway. (c) 30 top biological or disease pathways that were significantly enriched for genes that were up-regulated by at least 2-fold as a consequence of miR-574-5p exposure (Dotap-PS-miR-574-5p-treated versus Dotap-only treated, n = 3) as analyzed by the KEGG pathway analysis. The rich factor is the ratio between the number of the significantly-up-regulated genes mapped to the pathway and the total number of genes in the pathway and the Q-values were the p-values corrected with the false discovery rates