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. 2020 Sep 16;10(25):11535–11548. doi: 10.7150/thno.45261

Figure 3.

Figure 3

FOXO1(+) tumor cells promoted M2 macrophages recruitment by CCL20 secretion. (A) Relative expression of CCL20 in the FOXO1(+) and FOXO1(-) tumor cells detected using qRT-PCR (*P < 0.05; **P < 0.01). (B) Concentration of CCL20 in the supernatants of FOXO1(+) and FOXO1(-) tumor cells measured with ELISA (*P < 0.05; **P < 0.01). (C) Expression of protein CCL20 in FOXO1(+) and FOXO1(-) tumor cells detected with western blotting. (D) Expression of protein CCL20 in FOXO1(+) tumor cells and their FOXO1 silenced tumor cells detected with western blotting. (E) Representatives and summary of M2 macrophage migration assays induced with FOXO1(-) tumor cells, FOXO1(+) tumor cells, and FOXO1(+) tumor cells after blocking with 0.25 µg/mL and 1.0 µg/mL α-CCL20 antibody. Scale bar, 100 µm (*P < 0.05; **P < 0.01, ***P < 0.001). (F) Representatives and summary of M2 macrophage migration assays induced with serum-free medium, 5 ng/mL and 20 ng/mL CCL20 recombinant. Scale bar, 100 µm (*P < 0.05; **P < 0.01). (G) Representatives and summary of M2 macrophage migration assays induced with FOXO1(+)-Ctl tumor cells, FOXO1(+)-sh1 tumor cells, and FOXO1(+)-sh2 tumor cells. Scale bar, 100 µm (***P < 0.001; ****P < 0.0001).