Figure 6.

TNF-α expression is modulated by mmu-miR-181a-5p, while IL-10 gene expression is regulated by mmu-miR-21a-5p during Brucella abortus infection. First, we determined time dependent of mmu-miR-181a-5p (A) or mmu-miR-21a-5p (B) expression during B. abortus infection in bone marrow-derived macrophages (BMDMs). miRNAs expression was assessed by real-time PCR and was normalized to SNOR61. Error bars represent the mean ± SD. Similar results were obtained in two independent experiments. Statistically significant differences of miRNAs expression from infected BMDM compared with non-infected (NI) cells are denoted by an asterisk (p < 0.05). Second, mimics and inhibitors transfection for mmu-miR-181a-5p (C) or mmu-miR-21a-5p (D) showed increase or inhibition of specific miRNAs in BMDMs. Error bars represent the mean ± SD. Similar results were obtained in two independent experiments. Statistically significant differences of miRNAs availability from BMDM transfected with mimic to negative control to mimic (scramble) are denoted by an asterisk (p < 0.05), and # (p < 0.05) represents statistically significant differences between BMDMs transfected with anti-miR to negative control to inhibitors (scramble). Finally, BMDMs transfected with mimic or inhibitor for mmu-miR-21a-5p was infected with B. abortus and the level of IL-10 mRNA was measured by real-time PCR (E). BMDMs transfected with mimic or inhibitor for mmu-miR-181a-5p was infected with B. abortus and the level of TNF-α mRNA was determined by qPCR (F). The results were normalized to β-actin. Error bars represent the mean ± SD. Similar results were obtained in two independent experiments. Statistically significant differences of gene expression from infected cells to non-transfected (NT)/NI cells are denoted by an asterisk (p < 0.05), and # (p < 0.05) represents statistically significant differences with NT/infected (S2308) cells.