Figure 6
TNF-α mediates the induction of ISRE by activation of the NF-κB signaling.
(A) Confocal microscopy analysis of RelA induction and localization in Huh7 cells treated with TNF-α . RelA was induced and translocated to the nucleus upon TNF-α treatment. RelA antibody (green). Nuclei were visualized by DAPI (blue). (B) Western blot analysis confirmed the successful knockdown of RelA by lentiviral shRNA vectors in the Huh7 based ISRE luciferase reporter cells. (C) RelA knockdown largely blocked TNF-α induced ISRE-related luciferase activation as measured at 3 different time points (n = 3 independent experiments with 2–3 replicates each). (D) RelA knockdown largely blocked TNF-α induced ISG expression as measured by qRT-PCR (n = 4). (E) RelA knockdown has no significant influence on IFN-α induced ISRE-related luciferase activation as measured at 3 different time points (n = 3 independent experiments with 2–3 replicates each). Data presented as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant).
