Fig. 5.

Bleomycin enhances phospholipase D (PLD) activity, and PLD2 protein expression in bronchial airway epithelial (Beas2B) cells. Beas2B cells grown in 35-mm dishes (~90% confluence) were challenged with 10 mU bleomycin for 24 h. A: total PLD activity was measured by Amplex Red assay, as indicated in materials and methods. B: representative Western blot of PLD1, PLD2, and β-actin expression in total cell lysates from Beas2B cells of 3 independent experiments in triplicate. C: quantification of Western blots by densitometry/ImageJ analysis and fold changes were normalized to β-actin. D: primary human bronchial epithelial cells (HBEpCs) in 35-mm dishes were treated with bleomycin as in A, and total PLD activity was determined with and without bleomycin challenge, as indicated above. E: representative Western blot of PLD1, PLD2, and β-actin in total cell lysates from HBEpCs of 3 independent experiments in triplicate. F: quantification of Western blots by densitometry/ImageJ analysis and fold changes were normalized to β-actin. *P < 0.05, **P < 0.005.