Research Paper Volume 12, Issue 17 pp 16820—16836
Neutrophil extracellular traps amplify neutrophil recruitment and inflammation in neutrophilic asthma by stimulating the airway epithelial cells to activate the TLR4/ NF-κB pathway and secrete chemokines
Figure 6. Inhibition of the TLR4/NF-κB pathway reduced chemokine expression and neutrophil migration in vitro after stimulation of HBEs and A549 by NETs. (A) After treatment of HBEs and A549 cells with TAK-242 and NETs simultaneously, p-p65 NF-κB and p65 NF-κB expression was measured by western blot. (B and C) After treatment of HBEs and A549 cells with TAK-242 and NETs simultaneously, CXCL1, CXCL2, and CXCL8 levels in the supernatant were analyzed by Luminex and RT-qPCR. (D and E) After treatment of HBEs and A549 cells with TAK-242 and NETs simultaneously, the supernatant was collected and added into the lower chamber. The neutrophils were then cultured in the upper chamber and neutrophil migration was analyzed by transwell after cell culture supernatant was induced for 48 h (200X). #: P<0.05, ##: P<0.01, ###: P<0.001, ####: P<0.0001 vs control group. *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001 vs NA group. ns = no statistical difference vs NA or control group.