Figure 5
PVP-I inhibits TLR4 and MyD88 co-localization which were induced by inflammatory response. Airway epithelial cells were pre-treated with LPS (1000 ng/mL) and ATP (5 mM) and then exposed to 0.1% PVP-I. (a) The cells lysate were immunoprecipitated with anti- MyD88 and TLR4 antibody, and then immunoprecipitated proteins were detected using anti-TLR4 and anti-MyD88 antibodies. The pHNECs had the same experimental methods as the airway epithelial cells. PVP-I was found to inhibit the co-localization with TLR4 and MyD88 complex formation. Actin was used as a control for input. Airway epithelial cells were pre-treated with LPS and ATP for 60 min and then treated or not with PVP-I for 24 h. (b) Immunocytochemistry for TLR4 (red) and MyD88 (green) co-localization in A549 cells and pHNECs is shown. Increased expressions of TLR4 and MyD88 by LPS/ATP stimulation were inhibited by PVP-I treatment in A549 cells and pHNECs. The scale bar indicates 10 μm. Airway epithelial cells were pre-treated with LPS and ATP for 10 min and then treated or not with PVP-I for 10 min. (c) TLR4 and Myd88 colocalization was assessed by calculating the Pearson correlation coefficient on twenty images per condition using the JACoP plugin on ImageJ.
