Fig. 4

SPON2 activated IL10, CCL2 and CSF1 to enhance polarization to the M2 phenotype. a Schematic of the sample preparation for Q-RT-PCR and flow cytometry. b Q-RT-PCR showed the mRNA expression of M1 macrophage markers (Nos2 and Cd86) and M2 macrophage markers (Cd206 and Arg1). c Flow cytometric quantification showing the proportion of M2 macrophages (F4/80⁺, CD206⁺). d Correlation analysis of mRNA levels between SPON2 and cytokines that promote macrophage polarization in TCGA. Two-tailed Pearson test. n = 458. e Q-RT-PCR showed the mRNA expression of IL10, CCL2 and CSF1 in CRC cell lines. f Western blot analysis of IL10, CCL2 and CSF1 protein levels in the cell lysates of SW480/Vector, SW480/SPON2, SW620/Scramble, SW620/shSPON2#1 and SW620/shSPON2#2 cells. g Western blot analysis of CD206, MAC2 and CD68 protein levels in THP-1 cells cocultured with SW480/Vector, SW480/SPON2, SW620/Scramble, SW620/shSPON2#1 and SW620/shSPON2#2 cells. h-j ELISA analysis of IL10, CCL2 and CSF1 in SW480/Vector, SW480/SPON2, SW620/Scramble, SW620/shSPON2#1 and SW620/shSPON2#2 cells. In b, e, and h-j, data represent mean ± SD, Student’s t test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001