Fig. 3

DDX39B modulates NF-κB activity in association with LGP2. a Luciferase assay using the -1C reporter in wild-type, Rig-I^−/−, Mda5^−/−, and Lgp2^−/− MEFs following infection with sh-Ddx39b or a non-targeting control. Data show mean relative value, ± SEM of two independent experiments. b Immunoblot (IB) in the indicated MEFs infected with sh-Ddx39b or non-targeting control using anti-phospho-p65 antibody. c Luciferase assay in U87 cells stably expressing sh-DDX39B or non-targeting control transfected with si-LGP2 or si-control. Data show mean value, ± SEM of two independent experiments. d IB in A172 cells expressing sh-DDX39B or non-targeting control probed with the indicated antibodies. e Immunofluorescence (IF) staining (left) and quantification (right) of dsRNA in U87 DDX39B CRISPR cells. Data show mean value, ± SD of ten view fields, repeated. Scale bar, 10 μm. f Co-immunoprecipitation (Co-IP) in U87 cells transfected with empty vector or S-tagged DDX39B. IP with S-protein agarose followed by IB using the indicated antibodies. Input probed as shown. g Co-IP in U87 cells (left) and GL261 mouse GBM cells (right) following IP with anti-DDX39B or control IgG and IB with the indicated antibodies. Input probed as shown. h IF staining for endogenous DDX39B (FITC) in U87 cells. DAPI counter stain. Scale bar, 10 μm. Asterisk, cytoplasmic DDX39B; arrowhead, nuclear DDX39B. i Luciferase assay in Lgp2^−/− MEFs infected with sh-DDX39B shRNA or sh-control and transfected with empty vector (EV) or LGP2. Data show mean value, ± SEM of three independent experiments. j Luciferase assay in Lgp2^−/− MEFs cells infected with EV or S-tagged DDX39B and transfected with an independent EV or FLAG-LGP2. Data show mean value, ± SEM of three independent experiments. Inset: IB with anti-FLAG. *P < 0.05 (two-tailed t test)