Role of TcpF in NF-κB activation.

(A) The translocation of the p65 subunit of NF-κB to the nucleus of RAW264.7 macrophages infected (MOI = 10) with wild type, TcpF mutant (SPB03) and complemented strain (SPB04) for 1 hour or without infection (Control), was analyzed by subjecting the nuclear fraction to Western blot with p65 antibody. (B) The NF-κB transcriptional activity was assessed during infection of MEF cells by wild type, TcpF mutant and complemented strains. MEF cells were transfected with NF-κB-luciferase and Renilla-luciferase reporter constructs. After 24 hours, the medium was changed and cells were challenged with enterococci at a MOI of 10 or with LPS (0.5 µg/ml) as positive control. The data represent the mean values of three independent experiments and error bars indicate the standard deviations. Significance * P<0.05.