Figure 5: BjAnn1 attenuated the nuclear translocation of NF-κB and reduces its target gene expression. (A) Cytoplasmic and Nuclear lysates were prepared from U87 and U251 cells that are stably expressing control (EV) and BjAnn1 plasmids and stimulated with or without TNF-α for 30 min and subjected to Western blot analysis with p65 and p50 antibodies. GAPDH and Lamin B were used as loading controls for cytoplasmic and nuclear fractions respectively. (B) U87 and U251 cells stably expressing control (pEV) and BjAnn1 were transfected with the NF-κB-Luc reporter and after 48 h cells were stimulated with TNF-α; and the luciferase activity was measured. All data presented are the mean of 3 independent experiments with mean ± SE. *, p< 0.05, t test. (C) Whole cell lysates were prepared from U87 and U251 cells that are stably expressing control (EV) and BjAnn1 plasmids subjected to Western blot analysis with cyclin D1 and c-Myc antibodies. β-actin used as an loading control.