Figure 5

BAG and B. anthracis spores induce NF-κB activation. (A) RAW264.7 cells were cotransfected for 24 h with a firefly luciferase reporter plasmid regulated by the NF-κB transcription factor together with pRL-TK Renilla luciferase plasmid as an internal control for transfection efficiency. Then, the cells were stimulated with the indicated concentrations of BAG for a further 8 h. At the end of the stimulation period, the cells were lysed, and the dual luciferase activities were measured. Firefly luciferase activity was normalized to Renilla luciferase activity. Values are the mean ± SD of triplicate assays. *P <0.05 as compared with the untreated control group. (B) RAW264.7 cells were not stimulated or were stimulated with 1 μg/ml BAG or LPS or with B. anthracis spores (MOI of 10) for 1 h and then stained with anti-p65 followed by Alexa-488–conjugated secondary antibody and DAPI. Confocal images were obtained.