Fig. 5

Signaling pathways are involved in macrophage-induced cisplatin resistance and migration. (A) A549 cells were incubated in monocyte or macrophage CM for the indicated times. Phosphorylation or the expression level of each protein was detected by western blotting using specific antibodies. The experiment was repeated with similar results. (B) A549 cells were transfected with an NF-κB-dependent luciferase expression plasmid and then incubated in CM or treated with 15 ng/ml TNF-α for 8 h. (C) A549 cells were induced with macrophage CM in the presence or absence of each inhibitor for 24 h, respectively. Viable cells in each condition were measured and compared with the corresponding control. (D) A549 cells were incubated with macrophage CM containing PD98059 or IKK2 inhibitor IV (IKK2 i). Cisplatin was added into each cell for 8 h. Whole cell lysates were subjected to western blotting. (E, F) A549 cells were treated with macrophage CM containing each inhibitor for 24 h. Migration ability was determined using a trans migration assay and a wound healing assay. The wound healing assay was repeated six times and representative images are shown. Full-length blots are presented in Supplementary Fig. (2). Data are average values ± SD of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 (compared with Non-induced and vehicle)