Fig. 3

TBK1 forms aggregates at Golgi apparatus following potent RLR activation. a MEFs were either left untreated (MOCK) or transfected with HMW poly(I:C) (5 μg/mL) for 2 h (trPoly(I:C)). TBK1 staining was then analyzed by immunofluorescence. The Golgi apparatus was identified by labeling with an antibody raised against GM130. Scale bars, 10 μm. On the right, enlargement of the framed zone in the overlay. b WT or TBK1^–/– MEFs were stained with an antibody raised against TBK1, then analyzed by immunofluorescence analysis. c MEFs were either left untreated (MOCK) or transfected with HMW poly(I:C) (5 μg/mL) for 2 h (trPoly(I:C)). The indicated proteins were analyzed by immunofluorescence analysis with specific antibodies. Scale bars, 10 μm. d WT, MAVS^–/–, or STING^–/– MEFs were either left untreated (MOCK) or transfected with HMW poly(I:C) (5 μg/mL) for 4 h (trPoly(I:C)). TBK1 aggregation was assessed by immunofluorescence staining and aggregate counting. The data shown are means ± SD from three independent experiments (300 cells were counted per condition). ***P < 0.001 versus WT MEFs (Student’s t test). ns, not significant. e–g MEFs were either left untreated (MOCK) or transfected with HMW poly(I:C) (5 μg/mL) for 2 or 4 h (trPoly(I:C)). The indicated proteins were analyzed by immunofluorescence analysis with specific antibodies. The Golgi apparatus was stained with an antibody against GM130. Scale bars, 10 μm