Bleomycin could enhance the surface translocation of ANXA2 and activation of NF-κB pathway in lung cells

(A) A549 cells were treated with indicated concentrations of bleomycin (μg/ml) for 24 and 48 h, respectively. Western blot analysis of total expression and surface expression ANXA2 from EGTA elution (see Materials and methods) after bleomycin treatment (top). α-tubulin and β-actin were used as a control. The total p65 and phosphorylation status of p65 treated with or without bleomycin were measured by Western blot analysis (bottom). β-actin was used as an internal control. (B) Western blot analysis of total expression ANXA2 and surface expression ANXA2 from EGTA elution, as well as the total p65 and phosphorylation status of p65 in 16HBE cells treated with indicated concentrations of bleomycin (μg/ml) for 48 h. α-tublin or β-actin was used as a control. (C) Immunofluorescent detection of ANXA2 on the surface of A549 cells treated with or without 40 μg/ml bleomycin for 48 h. (D) Immunofluorescent detection of ANXA2 on the surface of 16HBE cells treated with or without 40 μg/ml bleomycin for 48 h. (E) Colocalization of ANXA2 with F-BLM on the surface of A549 cells. Microscope magnification: Σ 400×.