RNA-Seq analyses in miR-574-5p-transfected mouse splenocytes
RNA profiling by RNA-Seq analyses showed that in comparison with the control Dotap-treated cells, miR-574-5p transfection for 24 h resulted in significant up-regulation of 277 genes and down-regulation of 80 genes by at least 2-fold in the miR-574-5p-transfected splenocytes. Gene ontology and pathway enrichment analyses showed that the 277 significantly up-regulated genes in miR-574-5p-transfected cells were highly enriched and clustered in pathways associated with interferon and cytokine signaling, antiviral mechanisms, translesion synthesis and autoimmunity . Overall design: Mouse splenocytes were isolated from three 8-wk old female B6.WT mice with each sample divided in to two equivalent aliquots, one for Dotap-only and one for Dotap-PS-miR-574-5p transfection. Splenocytes were cultured in DMEM supplemented with 10% FBS, 2mM L-glutamine and 100 IU/ml penicillin and 100 µg/ml streptomycin for 4 h. Subsequently , cultured splenocytes were transfected with Dotap-only or Dotap-PS-miR-574-5p (Dotap-only-1, -2 and -3 and Dotap-miRNA-1, -2 and -3) at the concentration of 10 µg/ml. After 24 h of incubation, cells were harvested for RNA extraction and an alyses.