Figure 6

Effects of PBEF overexpression on IL-8 secretion and PBEF production in A549 cells. A. Representative western blotting image of PBEF and IL-8 protein detections. After starving for serum overnight, A549 cells were transfected with control (C), overexpressing construct pCAGGS-hPBEF (E) or vector pCAGGS (V) for 48 hours. Medium IL-8 and PBEF, Cell lysate PBEF and β-actin proteins were immunodetected as described in Figure 1 and Figure 2. B. Quantitation of PBEF and IL-8 protein levels by densitometric analysis. **p < 0.01 vs. control. C. representative western blotting image of PBEF and IL-8 protein detections with or without TNFα (15 ng/ml) treatment. A549 cells were transfected with control (C), pCAGGS-hPBEF (E) or vector pCAGGS (V) for 48 hours, and then stimulated by TNFα (15 ng/ml) for another 12 hours. Medium IL-8 and PBEF, cell lysate PBEF and β-actin proteins were immunodetected as described in above. D. quantitation of PBEF and IL-8 protein levels by densitometric analysis. **p < 0.01 vs. control.