Fig. 2

GCC2 knockdown decreases exosome release and abolishes the migration- and proliferation-promoting effects of cancer cell-derived exosomes. (A) Total number of exosomes released in control and GCC2 knockdown H460 cells were determined by nanoparticle tracking analysis. (B) Immunofluorescence results of CD63-positive exosomes (green) in H460 and H1299 cells. Cell nuclei were stained with Hoechst. Scale bar = 10 μm. H460 cell-derived, GCC2 + exosomes were collected and measured by nanoparticle tracking analysis, and H460 cells were treated with GCC2 + exosomes in a dose-dependent manner for 48 h. (C) Heat-map indicating the changes in mRNA expression levels of CSC- and EMT-related genes of H460 cells treated with GCC2 + exosomes. (D) Transwell migration and invasion assays were performed to assess the migratory ability of H460 cells treated with GCC2 + exosomes. Scale bar = 100 μm. (E) Simultaneously, colony formation assays were performed in H460 cells treated with GCC2 + exosomes. GCC2 knockdown H460 cell-derived, GCC2 KD exosomes were isolated and treated on H460 cells in a dose-dependent manner. (F) Heat-map indicating changes in mRNA expression levels of CSC- and EMT-related genes in H460 cells treated with GCC2 KD exosomes. (G) Transwell migration and invasion assays were performed to assess the migratory ability of H460 cells following GCC2 KD exosome treatment. Scale bar = 100 μm. (H) Colony formation abilities of H460 cells treated with GCC2 KD exosomes. (I) Graphs comparing results of migration, invasion, and colony formation assays of H460 cells treated with either GCC2 + or GCC2 KD exosomes in increasing doses. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.