Fig. 5.

Role of ubiquitin-specific peptidase 17 in the interaction between macrophages and lung cancer cells to enhance inflammation and stemness in cancer cells. a Schematic diagram of the macrophage recruitment assay. Conditioned medium collected from control and ubiquitin-specific peptidase 17 (USP17)-overexpressing cancer cells stimulated with or without 10 ng/ml interleukin-1β were added to the lower chamber of 0.4-μm transwell plates. Macrophages were plated on the upper chamber and incubated at 37 °C for 8 h. b Macrophages that migrated into the lower chamber were fixed, stained with 0.05% crystal violet, and counted. c Macrophages were cultured with conditioned media from control and USP17-overexpressing cells at 37 °C for 8 h. Inflammatory gene expressions in macrophages were analyzed with real-time quantitative polymerase chain reaction (RT-qPCR). d Schematic diagram of coculturing macrophages and cancer cells. The upper chamber of 0.4-μm transwell plates was cultured with or without macrophages; cancer cells were plated on the lower chamber. e, f The cells were incubated at 37 °C for 8 h. Stemness-associated (e) and inflammatory (f) gene expressions in the cancer cells were analyzed using RT-qPCR. Data represent mean ± standard deviation of three independent experiments, *P < 0.05; **P < 0.01 between the indicated groups, or compared with the control group