Fig. 3

Wnt5a induces M2 macrophages polarization via IL-10. a RT-qPCR analysis of the relative expression levels of M1 markers (HLA-DR, IL-12) and M2 markers (CD163, IL-10) in M0 macrophages treated with Wnt5a. b Flow cytometry analysis showed M2 polarization (CD163⁺) in M0 macrophages treated with Wnt5a. This M2 polarization was reversed by Box 5 (Wnt5a inhibitor). c RT-qPCR analysis of M2 polarization-associated cytokines expression in M0 macrophages with Wnt5a treatment. d Western blot analyzed the protein expression of IL-10 in Wnt5a-treated M0 macrophages. e ELISA analysis for IL10 secretion level in M0 macrophages treated with Wnt5a. f, g Flow cytometry analysis of M2 cells ratio in Wnt5a-stimulated M0 macrophages with or without IL-10 antibody or IgG or recombinant IL10 protein. h Confocal immunofluorescence analysis showed the co-localization expression of Wnt5a, CD163 and IL-10 in CRC sample. Bar = 25 μm. All experiments were performed independently at least three times. Statistical analysis was conducted using one-way ANOVA. Error bars, SEM. ns, not significant. *P < 0.05. **P < 0.01. ***P < 0.001