Figure 5

NF-κB was activated by the PKR protein-binding function in MIN6 cells. Twenty-four hours after transfection, MIN6 cells were treated with coumermycin as indicated. (A) Whole cell extracts (WCE) and prepared nuclear extracts (NE) were collected and analyzed by Western blotting with antibodies against IκBα and p65, respectively. (B) Administered as Figure 1B, p65 localizations in cytoplasm and nuclear of MIN6 cells were determined by IFA (scale bar = 100 µm). (C) Luciferase activity was measured and shown in fold. β-actin was detected as an internal control. Data are means ± SEM of three separate experiments. *P < 0.05 versus control.