Figure 3 YDZG inhibits LPS-induced NF-κB activation. THP-1 (A) or J774A.1 (B) cells were pretreated with the indicated concentrations of YDZG for 1 h before being stimulated with LPS (1 μg·mL-1) for another 2 h. The expression of IKK-α, IKK-β, p- IKK-α/β, IκB-α, p- IκB-α, NF-κB/p65, and p- NF-κB/p65 proteins was determined by Western blotting (n = 5). (C)THP-1 cells were pretreated with the indicated concentrations of YDZG for 1 h before being stimulated with LPS for another 2 h. p65 translocation was determined using the immunofluorescence assay described in the Methods section (n = 3). Primary antibody anti-NF-κB/p65 (1:100) and secondary antibody goat anti-Rabbit Alexa Fluor 568 (1:200) were employed (scale bar = 10 μm). (D) THP-1 cells were pretreated with YDZG (4 μM) for 1 h and then stimulated with LPS (1 μg·mL-1) for 1 h. The localization of p65 in the cytoplasm and nucleus was detected by Western blotting (n = 5).