The phenotype and cytokine secretion of polarized macrophages. PBMC-derived monocytes were used for macrophage induction and differentiation. PMA was used for the induction of macrophages, and TSN and B7-H3 were used for macrophage polarization. (A) Surface molecules (CD206 and HLA-DR) were examined by flow cytometry to identify the phenotype of macrophages. The figure presents overlaps of CD206/HLA-DR staining and control Ig staining. (B) Cytokines (IL-10 and TNF-α) were detected by ELISA, and (C) iNOS RNA expression was detected by reverse transcription-polymerase chain reaction to analyze the function of macrophages. All results together indicated that B7-H3 could promote M2 macrophage differentiation. CRC, colorectal cancer; B7-H3, B7 homolog 3 protein; PBMCs, peripheral blood mononuclear cells; TAMs, tumor-associated macrophages; PMA, phorbol 12-myristate 13-acetate; TSN, tumor supernatant; CD, cluster of differentiation; HLA-DR, human leukocyte antigen-antigen D related; IL-10, interleukin; TNF-α, tumor necrosis factor-α; iNOS, inducible nitric oxide synthase.