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. 2021 Feb 9;29(6):2088–2107. doi: 10.1016/j.ymthe.2021.02.006

Figure 1.

Figure 1

EMT-CRC cell-derived exosomes induce M2 macrophage polarization in vitro

(A) Schematic description of the experimental design. (B) Transmission electron microscopy images of exosomes isolated from the medium of HCT116 cells under normal (Nor-Exos) and EMT status (EMT-Exos). Scale bars, 100 nm. (C) Nanosight particle tracking analysis of Nor-Exos and EMT-Exos. (D) Western blot analysis of the expression of exosome markers (CD9, CD63, CD81, TSG101) in cells, Nor-Exos, and EMT-Exos. (E) Flow cytometry for analyzing the expression of HLA-DR, CD86, CD163, and CD206 in macrophages incubated with Nor/EMT-HT29-Exos or Nor/EMT-HCT116-Exos for 48 h. (F and G) qRT-PCR analyses of the macrophage-associated markers in macrophages cocultured with Nor/EMT-HT29-Exos or Nor/EMT-HCT116-Exos for 48 h. (H and I) ELISA for analyzing the secretion of IL-10 and IL-12 in Nor/EMT-HT29-Exos or Nor/EMT-HCT116-Exos incubated macrophages for 48 h. Experiments were performed in triplicates. Error bars, SEM. Statistical analysis was conducted using one-way ANOVA. ∗p < 0.05; ∗∗p < 0.01.