Fig. 4. MIB2 inhibits host innate immunity.

A Relative protein and mRNA levels of MIB2 in MSpC cells after transfected with si-NC or si-circMIB2. B Relative protein levels of TRAF6 in MSpC cells after transfected with si-NC or si-MIB2 and in MIC cells with pcDNA3.1 or MIB2. C, D qPCR assays were performed to determine the expression levels of antiviral genes in MSpC cells transfected with si-NC or si-MIB2 and vector or MIB2 after SCRV infection (C) and determine the expression levels of T inflammatory factors in MIC cells transfected with si-NC or si-MIB2 and pcDNA3.1 or MIB2 after V. anguillarum infection (D). E Relative luciferase activities were detected in MSpC after co-transfection with TRAF6 expression plasmid, phRL-TK Renilla luciferase plasmid, luciferase reporters, pcDNA3.1, MIB2. F MSpC cells transfected with NC or si-MIB2 and pcDNA3.1 or MIB2, respectively, then infected with SCRV. The qPCR analysis was conducted for SCRV RNA expression. G MIC cells were transfected with NC or si-MIB2 and pcDNA3.1 or MIB2 and then infected with V. anguillarum. H MSpC and MIC cells seeded in 48-well plates overnight were treated with cultural supernatants at the dose indicated for 48 h. Then, cell monolayers were fixed with 4% paraformaldehyde and stained with 1% crystal violet. MSpC cells were transfected with si-NC or si-MIB2; MIC cells were transfected with vector or MIB2. All data represented the mean ± SE from three independent triplicated experiments. *p < 0.05; **p < 0.01.