Figure 2

Dose-response and time-course of TNFα induced PBEF protein expression in A549 cells. A. Dose-response. The top panel is a typical western blotting image of PBEF and β-actin protein detections. After starving for serum overnight, A549 cells were stimulated with different doses of TNFα, as indicated, for 24 hours. Equal amount of total cell lysate protein from each sample was separated by 10% SDS-PAGE and immunodetected by the western blotting using anti-human PBEF or β-actin antibodies. Middle panel-Quantitation of secreted PBEF level by densitometric analysis. Results from each group are presented as mean ± SD of 4 samples from two separate experiments. The bottom panel is the quantitation of cell lysate PBEF protein level by densitometric analysis. B. Time-course. Top panel-Representative western blotting image of PBEF and β-actin protein detections. After starving for serum overnight, A549 cells were stimulated with TNFα (15 ng/ml) for different time (h) as indicated. Lower panel-Quantitation of cell lysate PBEF protein level by densitometric analysis. *p < 0.05 and **p < 0.01 vs 0 dosage of TNFα or 0 h control.