Figure 1 | Laboratory Investigation

Figure 1

From: GDF15 derived from both tumor-associated macrophages and esophageal squamous cell carcinomas contributes to tumor progression via Akt and Erk pathways

Figure 1

Peripheral blood monocyte (PBMo)-derived macrophages acquired M2-like characteristics by stimulation with conditioned medium of TE series ESCC cell line (TECM). (a) TECM exposure assay: PBMos were treated with 25 ng/ml recombinant human M-CSF for 6 days to induce macrophage-like differentiation, and then exposed to 50% TECM for 2 days. (b) IL-10 and IL-12 mRNA expression pattern as M2-like characteristics in PBMo-derived macrophages stimulated with TECM. Expression levels were determined by quantitative RT-PCR and compared with those of PBMo-derived macrophages, normalized to GAPDH expression. Results are mean±s.e.m. (n=4; *P<0.05). (c) Induction of M2 markers CD163 and CD204 mRNA in PBMo-derived macrophages stimulated with TECM. Expression levels were analyzed by quantitative RT-PCR. Results are mean±s.e.m. (n=4; *P<0.05). (d) Expression of CD163 and CD204 in PBMo-derived macrophages stimulated with TECM. Double immunofluorescence was performed using anti-CD163 or CD204 (red) plus macrophage marker anti-CD11b (green) in a TECM exposure assay. Coexpression of CD163 and CD204 was detectable in PBMo-derived macrophages stimulated with TECM. Nuclei were stained with DAPI (blue). Magnification × 40. Scale bar=20 μm. (e) Upregulation of protumorigenic factors VEGFA, MMP2, and MMP9. Expression levels were determined by quantitative RT-PCR. Results are mean±s.e.m. (n=4; *P<0.05).

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