TRK-Fused Gene (TFG), a protein involved in protein secretion pathways, is an essential component of the antiviral innate immune response
Fig 5
TFG is required for the activation of TBK1 and downstream signalling upon cytosolic RNA sensor activation.
(A) HeLa cells previously treated with non-targeting siRNA (-) or siRNA targeting TFG (siTFG #1) were left uninfected or infected with SeV for indicated time. Whole cell extracts (WCE) were used in immunoblot analysis with indicated antibodies. The same WCE were used in native-page under non-denaturing conditions to evaluate IRF3 dimerization. (B) MRC-5 fibroblasts were infected with different lentiviral vectors encoding different TFG-targeting shRNA (shTFG #1, 2 or 3) or a nontargeting (NT) control shRNA (shNT) and then subjected to puromycin selection as described in Materials and Methods. Cells were then left uninfected or infected with SeV for the times indicated. WCE were harvested and used in immunoblot analysis with indicated antibodies. β-actin was used as a loading control. These results are representative of at least three independent experiments with similar results. (C) WCE generated in (A) were used in immunoblot analysis with indicated antibodies. β-actin was used as a loading control. These results are representative of at least three independent experiments with similar results.