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. 2018 Jul 10;8:238. doi: 10.3389/fcimb.2018.00238

Figure 6.

Figure 6

TANK-binding kinase 1 (TBK1) attenuates Mycobacterium bovis (M. bovis) survival in vitro by control of autophagy and the production of interferon-β (IFN-β). (A) J774a.1 cells were treated with control or siTBK1 for 24 h and then infected with Mycobacterium bovis (M. bovis) at a multiplicity of infection (MOI) of 10 for 24 or 48 h. Quantitative analysis of the phosphorylated TBK1 (p-TBK1), IRF3 and phosphorylated IRF3 (p-IRF3) bands normalized to GAPDH is shown. (B) J774a.1 cells and (C) mouse bone marrow monocyte-derived macrophages (BMDMs) were treated with control or siTBK1 for 24 h, followed by M. bovis infection at a MOI of 10 for the indicated time. Production of IFN-β was measured by ELISA assay. (D) J774a.1 cells and (E) mouse bone marrow monocyte-derived macrophages (BMDMs) were treated with control, siTBK1 for 24 h, BX795 (5 μM, 2 h) and IFNAR1-blocking monoclonal antibody MAR1-5A3 (1 μg/ml, 2 h), and then infected with Mycobacterium bovis (M. bovis) at a multiplicity of infection (MOI) of 10 for 24 or 48 h. M. bovis survival was analyzed by colony-forming units (CFU). (F) Model depicting the cascade of events following M. bovis infection involving sequential upregulation of miR-199a; downregulation of TBK1; suppression of autophagosomes maturation; downregulation of IFN- β and increased survival of M. bovis. The red arrows show an inhibition of TBK1 by miR-199a seems high importance for the abrogation of autophagy than down regulation of IFN-β via TBK1/IRF3 axis. Data is presented as mean ± SD in three independent experiments (*P < 0.05; **P < 0.01).