Fig. 2

SPON2 derived from CRC cells promotes TAM migration and infiltration in tumors. a Migration of primary mouse macrophages (M0/M) toward conditioned medium from stable cell lines. Scale bar, 100 μm. b Left panel, representative immunofluorescence for MAC2 and F4/80 in growth factor-reduced Matrigel plugs supplemented with PBS or rSPON2 (1 mg/mL) and subcutaneously injected into C57 BL/6 mice. Right panel, quantification of F4/80⁺MAC2⁺ macrophages in Matrigel plugs. Scale bar, 100 μm. c Gross images of MC38/Scramble and MC38/shSpon2#1 subcutaneous tumors in C57 BL/6 mice. d Tumor growth curve of MC38/Scramble and MC38/shSpon2#1 subcutaneous tumors in C57 BL/6 mice. Two-way ANOVA test, *p < 0.05. e Weights of MC38/Scramble and MC38/shSpon2#1 subcutaneous tumors. Student’s t test, *p < 0.05. f Flow cytometric quantification showing the decreased infiltration of M2-TAMs (CD45⁺/CD11b⁺/F4/80⁺/CD206⁺). Student’s t test, *p < 0.05. g Representative images of MC38/Scramble and MC38/shSpon2#1 subcutaneous tumors. Hematoxylin and eosin (H&E) staining shows the histology of tumor tissues. h The representative gross images of the intestines and livers from MC38/Scramble and MC38/ shSpon2#1 orthotopic tumors. Sections of the liver were stained with H&E. Scale bar, 50 μm; the right graph shows quantification of no metastatic sample and liver metastatic sample. i Immunofluorescence (left panel) and quantification (right panel) of the macrophage marker MAC2 in MC38/Scramble and MC38/ shSpon2#1 orthotopic tumors. Scale bar, 100 μm. j Flow cytometric quantification showing the infiltration of M2-TAMs in orthotopic tumors. Data represent mean ± SD, ** p < 0.01, *** p < 0.001, **** p < 0.0001