Figure 5.

Gefitinib inhibits M2-like macrophage-promoted invasion and migration in vitro. (A) Schematic representation of the experimental approach used in vitro as described in the Methods section. BMDMs were treated with IL-13 (10 ng/mL) with or without gefitinib (0.62 μmol/L) for 72 h and the culture medium was replaced with fresh serum-free medium. After 24 h, the supernatant medium was collected as macrophage-conditioned medium (CM). (B) LLCs were cultured with CM for 24 h. Cell survival was determined by SRB assay. (C) For the wound healing assay, LLCs cells were seeded in 24-well plates in DMEM medium with 10% FBS, cells were then wounded with a straight scratch using a pipette tip and treated with CMs for 24 h. (D and E) The cells were seeded in the upper chamber and CM was placed into the lower chambers and allowed to incubate at 37 °C for 24 h. The migrated cells on the bottom chamber were stained with 0.1% crystal violet. Then, the effect of different CMs on RAW 264.7 and LLCs migration was evaluated by Transwell assay. The results are presented as the mean±SEM of triplicate determinations. ^*P<0.05.