<p>A498 cells were incubated in the absence or presence of physalin F (10 µg/mL) for indicated time and detection the expression of phospho-IκBα and IκBα (A). (B) The nuclear and cytosol extract were prepared as indicated in Methods and Materials and were analyzed by Western blotting for the detection of specific protein, as indicated (p-p65, p65, p50, GAPDH, and nucleolin). (C) cells were exposed to physalin F (10 µg/mL) for 6 hr and nuclear extracts were incubated with a hot NF-κB probe (lane 2–4) or cold probe (lane 1, indicate “cold”) and demonstrate the specificity of the bands obtained on EMSA. The data are representative of three independent experiments. P indicates positive nuclear extract. ** <i>P</i><0.01 compared with the control group. (D) Effect of ROS scavengers on physalin F-inhibited NF-κB nuclear translocation in A498 cells. Cells were pretreated with NAC (10 mM) and GSH (3 mM) for 0.5 hr and then treated with 10 µg/mL of physalin F for 6 hr. The nuclear extract were prepared as indicated in Methods and Materials. The levels of p65 and p50 were detected by Western blotting.</p
