Fig. 3

miR-3473 targets TRAF3. a The putative binding sites of miR-3473 in the TRAF3 3′-UTR are shown (TargetScanMouse 6.2, http://www.targetscreening.org/). b TRAF3 protein levels were measured by western blotting in Bp, Bt or uninfected macrophages at 24 h post infection. c TRAF3 mRNA levels were measured in Bp, Bt or uninfected macrophages by qRT-PCR at different time post infection (8 h, 12 h, 24 h) with actin as a reference gene. d The putative binding sites of miR-3473 in the TRAF3 3′-UTR were mutated as shown. e qRT-PCR analysis of miR-3473 expression in macrophages transfected with mimic, inhibitor, mimic control or inhibitor control oligonucleotides of miR-3473. f Macrophages treated with oligonucleotides (as Figure e) were co-transfected with a pMIR-Report-TRAF3-3′-UTR-luciferase or pMIR-Report-TRAF3-mut-3′-UTR-luciferase or pRL-TK vector as an internal control. After 24 h treatment, cell extractions were analyzed by luciferase reporter assay and values normalized to renilla activity. Data is represented as mean ± SEM from three independent experiments. *P value < 0.05, **P value < 0.01