However, flow cytometry analysis of macrophages co-cultured with tumor cells (MC38/Scramble, MC38/shSpon2#1 and MC38/shSpon2#2) showed that knockdown SPON2 significantly decreased the polarization toward M2-like macrophages (Supplementary Figure S3).​ TCGA-COAD data showed that the mRNA expression of SPON2 was positively correlated with the mRNA expression of M2 polarization factors such as CCL2, CSF1 and IL10 but not with M1 polarization factors, such as TNF-α, IFN-γ and IL12 (Fig. 4d). This evidence concerns the gene IFNG and neoplasm.