Figure 4

Effect of MME on the translocation and activation of NF-κB in LPS-stimulated BV-2 cells. (A) Cells were treated with and without MME for 2 h followed by LPS stimulation for 30 min. NF-κB/p65 subunits were probed by anti-NF-κB antibody and Alexa Fluor® 488-conjugated secondary antibody. The nuclei were stained by DAPI and the images were captured by confocal microscopy (×40). (B) Cells pretreated with different concentrations of MME for 2 h were stimulated with LPS for 30 min. Cytosolic and nuclear extracts were prepared and analyzed using Western blot by using corresponding antibodies. (C) Cells were co-transfected with 2 μg of NF-κB promoter-containing luciferase DNA along with 40 ng of control pRL-TK DNA for 40 h. Transfected cells were pretreated with various concentrations of MME for 2 h and then stimulated with LPS for 6 h. Cell lysates were prepared and used for reporter gene assay. Data are means ± SDs of three independent experiments. ^a p < 0.05 indicates significant differences compared to the non-treated group. ^b p < 0.05 indicates significant differences compared to the LPS-only group.