Fig. 2: miR-199b-5p and miR-2989 inhibit the humoral immunity pathway.

a Expression levels of humoral immunity genes, domeless (Dome) and toll-7 (Toll-7), were quantified by qPCR in SL1A cells 48 h after infection with SmBV. b Expression levels of domeless (Dome) and toll-7 (Toll-7) were quantified by qPCR in SL1A cells 48 h after transfection with mimic of miR-199b-5p and miR-2989, and a control miRNA, respectively. c Reporter plasmids pKShE-199b-5p 3′UTR and pKShE-2989 3′UTR in which the 3′-UTR of Domeless or Toll-7 were fused to the 3′-end of an EGFP coding region, respectively. These two reporter plasmids were transfected into SL1A cells with corresponding miRNA mimic or a control miRNA (Control). EGFP expression was detected at 48 h after transfection and quantified by d western blotting analysis and e fluorescent microscopy. Genes in JAK/STAT (f) and Toll (g) pathways decreased in expression after the transfection of miR-199b-5p- or miR-2989 mimic. The expression levels of genes in the signal cascade of the JAK/STAT pathway [domeless (DOME), STAT (STAT), and TEP (TEP)] and the Toll pathway [toll (TOLL), myd88 (MYD88), pelle (PELLE), dif (DIF), and drosomycin (DRO)] were determined by qPCR in SL1A cells transfected with miRNA mimics of miR-199b-5p and miR-2989, respectively. An actin signal was used as an internal control to normalize all readings. All experiments were performed with three biological replicates (n = 3). Data are expressed as the mean and standard deviation (SD). p-value were calculated using Student’s t-test (*p < 0.05).